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More progress on Ch2 results

Ryan C. Thompson 5 年之前
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共有 1 个文件被更改,包括 744 次插入128 次删除
  1. 744 128
      thesis.lyx

+ 744 - 128
thesis.lyx

@@ -226,7 +226,8 @@ LatexCommand tableofcontents
 status open
 status open
 
 
 \begin_layout Plain Layout
 \begin_layout Plain Layout
-Look into auto-generated nomenclature list: https://wiki.lyx.org/Tips/Nomenclature
+Look into auto-generated nomenclature list: https://wiki.lyx.org/Tips/Nomenclature.
+ Otherwise, do a manual pass for all abbreviations.
 \end_layout
 \end_layout
 
 
 \end_inset
 \end_inset
@@ -263,6 +264,36 @@ Search and replace: naive -> naïve
 \end_inset
 \end_inset
 
 
 
 
+\end_layout
+
+\begin_layout Standard
+\begin_inset Flex TODO Note (inline)
+status open
+
+\begin_layout Plain Layout
+Once all 3 content chapters are written, go back over them and make them
+ consistent in terms of 
+\begin_inset Quotes eld
+\end_inset
+
+we did X
+\begin_inset Quotes erd
+\end_inset
+
+ vs 
+\begin_inset Quotes eld
+\end_inset
+
+X was done
+\begin_inset Quotes erd
+\end_inset
+
+.
+\end_layout
+
+\end_inset
+
+
 \end_layout
 \end_layout
 
 
 \begin_layout Chapter*
 \begin_layout Chapter*
@@ -550,7 +581,7 @@ Chapter author list: Me, Sarah, Dan
 status open
 status open
 
 
 \begin_layout Plain Layout
 \begin_layout Plain Layout
-Need better section titles throughout the chapter
+Need better section titles throughout the entire chapter
 \end_layout
 \end_layout
 
 
 \end_inset
 \end_inset
@@ -3465,25 +3496,11 @@ effective promoter radii
 \begin_inset Quotes erd
 \begin_inset Quotes erd
 \end_inset
 \end_inset
 
 
- were used to define the promoter regions for all further analyses.
-\end_layout
-
-\begin_layout Standard
-\begin_inset Flex TODO Note (inline)
-status open
-
-\begin_layout Plain Layout
-Clarify that radius depends on histone mark but 
-\emph on
-not
-\emph default
- experimental condition.
- 
-\end_layout
-
-\end_inset
-
-
+ remain approximately the same across all combinations of experimental condition
+ (cell type, time point, and donor), so they appear to be a property of
+ the histone mark itself.
+ Hence, these radii were used to define the promoter regions for each histone
+ mark in all further analyses.
 \end_layout
 \end_layout
 
 
 \begin_layout Standard
 \begin_layout Standard
@@ -3511,6 +3528,20 @@ wide false
 sideways false
 sideways false
 status open
 status open
 
 
+\begin_layout Plain Layout
+\begin_inset Flex TODO Note (inline)
+status open
+
+\begin_layout Plain Layout
+This figure is generated from the old analysis.
+ Eiher note that in some way or re-generate it from the new peak calls.
+\end_layout
+
+\end_inset
+
+
+\end_layout
+
 \begin_layout Plain Layout
 \begin_layout Plain Layout
 \align center
 \align center
 \begin_inset Graphics
 \begin_inset Graphics
@@ -3543,18 +3574,53 @@ Expression distributions of genes with and without promoter peaks.
 
 
 \end_layout
 \end_layout
 
 
-\begin_layout Plain Layout
+\end_inset
+
 
 
 \end_layout
 \end_layout
 
 
+\begin_layout Standard
+H3K4me2 and H3K4me2 have previously been reported as activating marks, while
+ H3K27me3 has been reported as inactivating [CITE].
+ The data are consistent with this characterization: genes whose promoters
+ (as defined by the radii for each histone mark described above) overlap
+ with a H3K4me2 or H3K4me3 peak tend to have higher expression than those
+ that don't, while H3K27me3 is likewise associated with lower gene expression,
+ as shown in 
+\begin_inset CommandInset ref
+LatexCommand ref
+reference "fig:fpkm-by-peak"
+plural "false"
+caps "false"
+noprefix "false"
+
+\end_inset
+
+.
+ This pattern holds across all combinations of cell type and time point
+ (Welch's 
+\emph on
+t
+\emph default
+-test, all 
+\begin_inset Formula $p\mathrm{-values}\ll2.2\times10^{-16}$
 \end_inset
 \end_inset
 
 
+).
+ The difference in average FPKM values when a peak overlaps the promoter
+ is about 
+\begin_inset Formula $+5.67$
+\end_inset
 
 
-\end_layout
+ for H3K4me2, 
+\begin_inset Formula $+5.76$
+\end_inset
 
 
-\begin_layout Itemize
-H3K4 is correlated with higher expression, and H3K27 is correlated with
- lower expression genome-wide
+ for H3K4me2, and 
+\begin_inset Formula $-4.00$
+\end_inset
+
+ for H3K27me3.
 \end_layout
 \end_layout
 
 
 \begin_layout Standard
 \begin_layout Standard
@@ -3562,9 +3628,10 @@ H3K4 is correlated with higher expression, and H3K27 is correlated with
 status open
 status open
 
 
 \begin_layout Plain Layout
 \begin_layout Plain Layout
-Grr, gotta find these figures.
- Maybe in the old analysis? At least one of these plots is definitely in
- Sarah's paper.
+I also have some figures looking at interactions between marks (e.g.
+ what if a promoter has both H3K4me3 and H3K27me3), but I don't know if
+ that much detail is warranted here, since all the effects just seem approximate
+ly additive anyway.
 \end_layout
 \end_layout
 
 
 \end_inset
 \end_inset
@@ -3572,25 +3639,11 @@ Grr, gotta find these figures.
 
 
 \end_layout
 \end_layout
 
 
-\begin_layout Itemize
-Figures showing these correlations: box/violin plots of expression distributions
- with every combination of peak presence/absence in promoter
-\end_layout
-
-\begin_layout Itemize
-Appropriate statistical tests showing significant differences in expected
- directions
-\end_layout
-
 \begin_layout Subsection
 \begin_layout Subsection
 RNA-seq and H3K4 methylation patterns in naive and memory show convergence
 RNA-seq and H3K4 methylation patterns in naive and memory show convergence
  at day 14
  at day 14
 \end_layout
 \end_layout
 
 
-\begin_layout Standard
-
-\end_layout
-
 \begin_layout Standard
 \begin_layout Standard
 \begin_inset ERT
 \begin_inset ERT
 status open
 status open
@@ -4316,7 +4369,10 @@ Check up on figure refs in this paragraph
 \end_layout
 \end_layout
 
 
 \begin_layout Standard
 \begin_layout Standard
-Figure 
+We hypothesized that if naive cells had differentiated into memory cells
+ by Day 14, then their patterns of expression and histone modification should
+ converge with those of memory cells at Day 14.
+ Figure 
 \begin_inset CommandInset ref
 \begin_inset CommandInset ref
 LatexCommand ref
 LatexCommand ref
 reference "fig:PCoA-promoters"
 reference "fig:PCoA-promoters"
@@ -4374,8 +4430,10 @@ noprefix "false"
 \end_inset
 \end_inset
 
 
 ), which accounts for shared variation across all 3 histone marks and the
 ), which accounts for shared variation across all 3 histone marks and the
- RNA-seq data, confirming that this is a coordinated pattern across all
- 4 data sets.
+ RNA-seq data, confirming that this convergence is a coordinated pattern
+ across all 4 data sets.
+ While this observation does not prove that the naive cells have differentiated
+ into memory cells at Day 14, it is consistent with that hypothesis.
 \end_layout
 \end_layout
 
 
 \begin_layout Subsection
 \begin_layout Subsection
@@ -4386,10 +4444,25 @@ Effect of promoter coverage upstream vs downstream of TSS
 \begin_inset Flex TODO Note (inline)
 \begin_inset Flex TODO Note (inline)
 status open
 status open
 
 
+\begin_layout Plain Layout
+There is enough here for multiple sections.
+ At least one each for H3K4me2 and H3K27me3.
+\end_layout
+
+\end_inset
+
+
+\end_layout
+
+\begin_layout Standard
+\begin_inset Flex TODO Note (inline)
+status open
+
 \begin_layout Plain Layout
 \begin_layout Plain Layout
 For the figures in this section, the group labels are arbitrary, so if time
 For the figures in this section, the group labels are arbitrary, so if time
  allows, it would be good to manually reorder them in a logical way, e.g.
  allows, it would be good to manually reorder them in a logical way, e.g.
  most upstream to most downstream.
  most upstream to most downstream.
+ If this is done, make sure to update the text with the correct group labels.
 \end_layout
 \end_layout
 
 
 \end_inset
 \end_inset
@@ -4424,7 +4497,7 @@ begin{landscape}
 \begin_inset Float figure
 \begin_inset Float figure
 wide false
 wide false
 sideways false
 sideways false
-status collapsed
+status open
 
 
 \begin_layout Plain Layout
 \begin_layout Plain Layout
 \align center
 \align center
@@ -4476,7 +4549,7 @@ Average relative coverage for each bin in each cluster
 \begin_inset Float figure
 \begin_inset Float figure
 wide false
 wide false
 sideways false
 sideways false
-status collapsed
+status open
 
 
 \begin_layout Plain Layout
 \begin_layout Plain Layout
 \align center
 \align center
@@ -4522,7 +4595,7 @@ PCA of relative coverage depth, colored by K-means cluster membership.
 \begin_inset Float figure
 \begin_inset Float figure
 wide false
 wide false
 sideways false
 sideways false
-status collapsed
+status open
 
 
 \begin_layout Plain Layout
 \begin_layout Plain Layout
 \align center
 \align center
@@ -4568,6 +4641,12 @@ Gene expression grouped by promoter coverage clusters.
 \begin_layout Plain Layout
 \begin_layout Plain Layout
 
 
 \series bold
 \series bold
+\begin_inset CommandInset label
+LatexCommand label
+name "fig:H3K4me2-neighborhood"
+
+\end_inset
+
 K-means clustering of promoter H3K4me2 relative coverage depth in naive
 K-means clustering of promoter H3K4me2 relative coverage depth in naive
  day 0 samples.
  day 0 samples.
  
  
@@ -4629,44 +4708,19 @@ shape
 \end_layout
 \end_layout
 
 
 \begin_layout Standard
 \begin_layout Standard
-\begin_inset Float figure
-wide false
-sideways false
-status collapsed
-
-\begin_layout Plain Layout
-\align center
-\begin_inset Float figure
-wide false
-sideways false
-status collapsed
+\begin_inset ERT
+status open
 
 
 \begin_layout Plain Layout
 \begin_layout Plain Layout
-\align center
-\begin_inset Graphics
-	filename graphics/CD4-csaw/ChIP-seq/H3K27me3-neighborhood-clusters-CROP.png
-	lyxscale 25
-	width 30col%
-	groupId covprof-subfig
-
-\end_inset
 
 
 
 
+\backslash
+end{landscape}
 \end_layout
 \end_layout
 
 
-\begin_layout Plain Layout
-\begin_inset Caption Standard
-
 \begin_layout Plain Layout
 \begin_layout Plain Layout
 
 
-\series bold
-\begin_inset CommandInset label
-LatexCommand label
-name "fig:H3K27me3-neighborhood-clusters"
-
-\end_inset
-
-Average relative coverage for each bin in each cluster
+}
 \end_layout
 \end_layout
 
 
 \end_inset
 \end_inset
@@ -4674,107 +4728,669 @@ Average relative coverage for each bin in each cluster
 
 
 \end_layout
 \end_layout
 
 
+\begin_layout Standard
+To test whether the position of a histone mark relative to a gene's transcriptio
+n start site (TSS) was important, we looked at the 
+\begin_inset Quotes eld
 \end_inset
 \end_inset
 
 
-
-\begin_inset space \hfill{}
+landscape
+\begin_inset Quotes erd
 \end_inset
 \end_inset
 
 
+ of ChIP-seq read coverage in naive Day 0 samples within 5 kb of each gene's
+ TSS by binning reads into 500-bp windows tiled across each promoter LogCPM
+ values were calculated for the bins in each promoter and then the average
+ logCPM for each promoter's bins was normalized to zero, such that the values
+ represent coverage relative to other regions of the same promoter rather
+ than being proportional to absolute read count.
+ The promoters were then clustered based on the normalized bin abundances
+ using 
+\begin_inset Formula $k$
+\end_inset
 
 
-\begin_inset Float figure
-wide false
-sideways false
-status collapsed
-
-\begin_layout Plain Layout
-\align center
-\begin_inset Graphics
-	filename graphics/CD4-csaw/ChIP-seq/H3K27me3-neighborhood-PCA-CROP.png
-	lyxscale 25
-	width 30col%
-	groupId covprof-subfig
-
+-means clustering with 
+\begin_inset Formula $K=6$
 \end_inset
 \end_inset
 
 
+.
+ Different values of 
+\begin_inset Formula $K$
+\end_inset
 
 
+ were also tested, but did not substantially change the interpretation of
+ the data.
 \end_layout
 \end_layout
 
 
-\begin_layout Plain Layout
-\begin_inset Caption Standard
+\begin_layout Standard
+For H3K4me2, plotting the average bin abundances for each cluster reveals
+ a simple pattern (Figure 
+\begin_inset CommandInset ref
+LatexCommand ref
+reference "fig:H3K4me2-neighborhood-clusters"
+plural "false"
+caps "false"
+noprefix "false"
 
 
-\begin_layout Plain Layout
+\end_inset
 
 
-\series bold
-\begin_inset CommandInset label
-LatexCommand label
-name "fig:H3K27me3-neighborhood-pca"
+): Cluster 5 represents a completely flat promoter coverage profile, likely
+ consisting of genes with no H3K4me2 methylation in the promoter.
+ All the other clusters represent a continuum of peak positions relative
+ to the TSS.
+ In order from must upstream to most downstream, they are Clusters 6, 4,
+ 3, 1, and 2.
+ There do not appear to be any clusters representing coverage patterns other
+ than lone peaks, such as coverage troughs or double peaks.
+ Next, all promoters were plotted in a PCA plot based on the same relative
+ bin abundance data, and colored based on cluster membership (Figure 
+\begin_inset CommandInset ref
+LatexCommand ref
+reference "fig:H3K4me2-neighborhood-pca"
+plural "false"
+caps "false"
+noprefix "false"
 
 
 \end_inset
 \end_inset
 
 
-PCA of relative coverage depth, colored by K-means cluster membership.
-\end_layout
+).
+ The PCA plot shows Cluster 5 (the 
+\begin_inset Quotes eld
+\end_inset
 
 
+no peak
+\begin_inset Quotes erd
 \end_inset
 \end_inset
 
 
+ cluster) at the center, with the other clusters arranged in a counter-clockwise
+ arc around it in the order noted above, from most upstream peak to most
+ downstream.
+ Notably, the 
+\begin_inset Quotes eld
+\end_inset
 
 
-\end_layout
+clusters
+\begin_inset Quotes erd
+\end_inset
 
 
+ form a single large 
+\begin_inset Quotes eld
 \end_inset
 \end_inset
 
 
+cloud
+\begin_inset Quotes erd
+\end_inset
 
 
-\begin_inset space \hfill{}
+ with no apparent separation between them, further supporting the conclusion
+ that these clusters represent an arbitrary partitioning of a continuous
+ distribution of promoter coverage landscapes.
+ While the clusters are a useful abstraction that aids in visualization,
+ they are ultimately not an accurate representation of the data.
+ A better representation might be something like a polar coordinate system
+ with the origin at the center of Cluster 5, where the radius represents
+ the peak height above the background and the angle represents the peak's
+ position upstream or downstream of the TSS.
+ The continuous nature of the distribution also explains why different values
+ of 
+\begin_inset Formula $K$
 \end_inset
 \end_inset
 
 
+ led to similar conclusions.
+\end_layout
 
 
-\begin_inset Float figure
-wide false
-sideways false
-status collapsed
+\begin_layout Standard
+\begin_inset Flex TODO Note (inline)
+status open
 
 
 \begin_layout Plain Layout
 \begin_layout Plain Layout
-\align center
-\begin_inset Graphics
-	filename graphics/CD4-csaw/ChIP-seq/H3K27me3-neighborhood-expression-CROP.png
-	lyxscale 25
-	width 30col%
-	groupId covprof-subfig
+RNA-seq values in the plots use logCPM but should really use logFPKM or
+ logTPM.
+\end_layout
 
 
 \end_inset
 \end_inset
 
 
 
 
 \end_layout
 \end_layout
 
 
-\begin_layout Plain Layout
-\begin_inset Caption Standard
+\begin_layout Standard
+\begin_inset Flex TODO Note (inline)
+status open
 
 
 \begin_layout Plain Layout
 \begin_layout Plain Layout
-
-\series bold
-\begin_inset CommandInset label
-LatexCommand label
-name "fig:H3K27me3-neighborhood-expression"
+Should have a table of p-values on difference of means between Cluster 5
+ and the others.
+\end_layout
 
 
 \end_inset
 \end_inset
 
 
-Gene expression grouped by promoter coverage clusters.
+
 \end_layout
 \end_layout
 
 
+\begin_layout Standard
+To investigate the association between relative peak position and gene expressio
+n, we plotted the Naive Day 0 expression for the genes in each cluster (Figure
+ 
+\begin_inset CommandInset ref
+LatexCommand ref
+reference "fig:H3K4me2-neighborhood-expression"
+plural "false"
+caps "false"
+noprefix "false"
+
 \end_inset
 \end_inset
 
 
+).
+ Most genes in Cluster 5, the 
+\begin_inset Quotes eld
+\end_inset
 
 
-\end_layout
+no peak
+\begin_inset Quotes erd
+\end_inset
 
 
+ cluster, have low expression values.
+ Taking this as the 
+\begin_inset Quotes eld
 \end_inset
 \end_inset
 
 
+baseline
+\begin_inset Quotes erd
+\end_inset
 
 
-\end_layout
+ distribution when no H3K4me2 methylation is present, we can compare the
+ other clusters' distributions to determine which peak positions are associated
+ with elevated expression.
+ As might be expected, the 3 clusters representing peaks closest to the
+ TSS, Clusters 1, 3, and 4, show the highest average expression distributions.
+ Specifically, these clusters all have their highest ChIP-seq abundance
+ within 1kb of the TSS, consistent with the previously determined promoter
+ radius.
+ In contrast, cluster 6, which represents peaks several kb upstream of the
+ TSS, shows a slightly higher average expression than baseline, while Cluster
+ 2, which represents peaks several kb downstream, doesn't appear to show
+ any appreciable difference.
+ Interestingly, the cluster with the highest average expression is Cluster
+ 1, which represents peaks about 1 kb downstream of the TSS, rather than
+ Cluster 3, which represents peaks centered directly at the TSS.
+ This suggests that conceptualizing the promoter as a region centered on
+ the TSS with a certain 
+\begin_inset Quotes eld
+\end_inset
 
 
-\begin_layout Plain Layout
-\begin_inset Caption Standard
+radius
+\begin_inset Quotes erd
+\end_inset
 
 
-\begin_layout Plain Layout
+ may be an oversimplification – a peak that is a specific distance from
+ the TSS may have a different degree of influence depending on whether it
+ is upstream or downstream of the TSS.
+\end_layout
+
+\begin_layout Standard
+\begin_inset ERT
+status open
+
+\begin_layout Plain Layout
+
+
+\backslash
+afterpage{
+\end_layout
+
+\begin_layout Plain Layout
+
+
+\backslash
+begin{landscape}
+\end_layout
+
+\end_inset
+
+
+\end_layout
+
+\begin_layout Standard
+\begin_inset Float figure
+wide false
+sideways false
+status open
+
+\begin_layout Plain Layout
+\align center
+\begin_inset Float figure
+wide false
+sideways false
+status open
+
+\begin_layout Plain Layout
+\align center
+\begin_inset Graphics
+	filename graphics/CD4-csaw/ChIP-seq/H3K4me3-neighborhood-clusters-CROP.png
+	lyxscale 25
+	width 30col%
+	groupId covprof-subfig
+
+\end_inset
+
+
+\end_layout
+
+\begin_layout Plain Layout
+\begin_inset Caption Standard
+
+\begin_layout Plain Layout
+
+\series bold
+\begin_inset CommandInset label
+LatexCommand label
+name "fig:H3K4me3-neighborhood-clusters"
+
+\end_inset
+
+Average relative coverage for each bin in each cluster
+\end_layout
+
+\end_inset
+
+
+\end_layout
+
+\end_inset
+
+
+\begin_inset space \hfill{}
+\end_inset
+
+
+\begin_inset Float figure
+wide false
+sideways false
+status open
+
+\begin_layout Plain Layout
+\align center
+\begin_inset Graphics
+	filename graphics/CD4-csaw/ChIP-seq/H3K4me3-neighborhood-PCA-CROP.png
+	lyxscale 25
+	width 30col%
+	groupId covprof-subfig
+
+\end_inset
+
+
+\end_layout
+
+\begin_layout Plain Layout
+\begin_inset Caption Standard
+
+\begin_layout Plain Layout
+
+\series bold
+\begin_inset CommandInset label
+LatexCommand label
+name "fig:H3K4me3-neighborhood-pca"
+
+\end_inset
+
+PCA of relative coverage depth, colored by K-means cluster membership.
+\end_layout
+
+\end_inset
+
+
+\end_layout
+
+\end_inset
+
+
+\begin_inset space \hfill{}
+\end_inset
+
+
+\begin_inset Float figure
+wide false
+sideways false
+status open
+
+\begin_layout Plain Layout
+\align center
+\begin_inset Graphics
+	filename graphics/CD4-csaw/ChIP-seq/H3K4me3-neighborhood-expression-CROP.png
+	lyxscale 25
+	width 30col%
+	groupId covprof-subfig
+
+\end_inset
+
+
+\end_layout
+
+\begin_layout Plain Layout
+\begin_inset Caption Standard
+
+\begin_layout Plain Layout
+
+\series bold
+\begin_inset CommandInset label
+LatexCommand label
+name "fig:H3K4me3-neighborhood-expression"
+
+\end_inset
+
+Gene expression grouped by promoter coverage clusters.
+\end_layout
+
+\end_inset
+
+
+\end_layout
+
+\end_inset
+
+
+\end_layout
+
+\begin_layout Plain Layout
+\begin_inset Caption Standard
+
+\begin_layout Plain Layout
+
+\series bold
+\begin_inset CommandInset label
+LatexCommand label
+name "fig:H3K4me3-neighborhood"
+
+\end_inset
+
+K-means clustering of promoter H3K4me3 relative coverage depth in naive
+ day 0 samples.
+ 
+\series default
+H3K4me2 ChIP-seq reads were binned into 500-bp windows tiled across each
+ promoter from 5
+\begin_inset space ~
+\end_inset
+
+kbp upstream to 5
+\begin_inset space ~
+\end_inset
+
+kbp downstream, and the logCPM values were normalized within each promoter
+ to an average of 0, yielding relative coverage depths.
+ These were then grouped using K-means clustering with 
+\begin_inset Formula $K=6$
+\end_inset
+
+,
+\series bold
+ 
+\series default
+and the average bin values were plotted for each cluster (a).
+ The 
+\begin_inset Formula $x$
+\end_inset
+
+-axis is the genomic coordinate of each bin relative to the the transcription
+ start site, and the 
+\begin_inset Formula $y$
+\end_inset
+
+-axis is the mean relative coverage depth of that bin across all promoters
+ in the cluster.
+ Each line represents the average 
+\begin_inset Quotes eld
+\end_inset
+
+shape
+\begin_inset Quotes erd
+\end_inset
+
+ of the promoter coverage for promoters in that cluster.
+ PCA was performed on the same data, and the first two principal components
+ were plotted, coloring each point by its K-means cluster identity (b).
+ For each cluster, the distribution of gene expression values was plotted
+ (c).
+\end_layout
+
+\end_inset
+
+
+\end_layout
+
+\end_inset
+
+
+\end_layout
+
+\begin_layout Standard
+\begin_inset ERT
+status open
+
+\begin_layout Plain Layout
+
+
+\backslash
+end{landscape}
+\end_layout
+
+\begin_layout Plain Layout
+
+}
+\end_layout
+
+\end_inset
+
+
+\end_layout
+
+\begin_layout Standard
+\begin_inset Flex TODO Note (inline)
+status open
+
+\begin_layout Plain Layout
+Is there more to say here?
+\end_layout
+
+\end_inset
+
+
+\end_layout
+
+\begin_layout Standard
+All observations described above for H3K4me2 ChIP-seq also appear to hold
+ for H3K4me3 as well (Figure 
+\begin_inset CommandInset ref
+LatexCommand ref
+reference "fig:H3K4me3-neighborhood"
+plural "false"
+caps "false"
+noprefix "false"
+
+\end_inset
+
+).
+\end_layout
+
+\begin_layout Subsection
+Promoter coverage H3K27me3
+\end_layout
+
+\begin_layout Standard
+\begin_inset ERT
+status open
+
+\begin_layout Plain Layout
+
+
+\backslash
+afterpage{
+\end_layout
+
+\begin_layout Plain Layout
+
+
+\backslash
+begin{landscape}
+\end_layout
+
+\end_inset
+
+
+\end_layout
+
+\begin_layout Standard
+\begin_inset Float figure
+wide false
+sideways false
+status collapsed
+
+\begin_layout Plain Layout
+\align center
+\begin_inset Float figure
+wide false
+sideways false
+status collapsed
+
+\begin_layout Plain Layout
+\align center
+\begin_inset Graphics
+	filename graphics/CD4-csaw/ChIP-seq/H3K27me3-neighborhood-clusters-CROP.png
+	lyxscale 25
+	width 30col%
+	groupId covprof-subfig
+
+\end_inset
+
+
+\end_layout
+
+\begin_layout Plain Layout
+\begin_inset Caption Standard
+
+\begin_layout Plain Layout
 
 
 \series bold
 \series bold
+\begin_inset CommandInset label
+LatexCommand label
+name "fig:H3K27me3-neighborhood-clusters"
+
+\end_inset
+
+Average relative coverage for each bin in each cluster
+\end_layout
+
+\end_inset
+
+
+\end_layout
+
+\end_inset
+
+
+\begin_inset space \hfill{}
+\end_inset
+
+
+\begin_inset Float figure
+wide false
+sideways false
+status collapsed
+
+\begin_layout Plain Layout
+\align center
+\begin_inset Graphics
+	filename graphics/CD4-csaw/ChIP-seq/H3K27me3-neighborhood-PCA-CROP.png
+	lyxscale 25
+	width 30col%
+	groupId covprof-subfig
+
+\end_inset
+
+
+\end_layout
+
+\begin_layout Plain Layout
+\begin_inset Caption Standard
+
+\begin_layout Plain Layout
+
+\series bold
+\begin_inset CommandInset label
+LatexCommand label
+name "fig:H3K27me3-neighborhood-pca"
+
+\end_inset
+
+PCA of relative coverage depth, colored by K-means cluster membership.
+\end_layout
+
+\end_inset
+
+
+\end_layout
+
+\end_inset
+
+
+\begin_inset space \hfill{}
+\end_inset
+
+
+\begin_inset Float figure
+wide false
+sideways false
+status collapsed
+
+\begin_layout Plain Layout
+\align center
+\begin_inset Graphics
+	filename graphics/CD4-csaw/ChIP-seq/H3K27me3-neighborhood-expression-CROP.png
+	lyxscale 25
+	width 30col%
+	groupId covprof-subfig
+
+\end_inset
+
+
+\end_layout
+
+\begin_layout Plain Layout
+\begin_inset Caption Standard
+
+\begin_layout Plain Layout
+
+\series bold
+\begin_inset CommandInset label
+LatexCommand label
+name "fig:H3K27me3-neighborhood-expression"
+
+\end_inset
+
+Gene expression grouped by promoter coverage clusters.
+\end_layout
+
+\end_inset
+
+
+\end_layout
+
+\end_inset
+
+
+\end_layout
+
+\begin_layout Plain Layout
+\begin_inset Caption Standard
+
+\begin_layout Plain Layout
+
+\series bold
+\begin_inset CommandInset label
+LatexCommand label
+name "fig:H3K27me3-neighborhood"
+
+\end_inset
+
 K-means clustering of promoter H3K27me3 relative coverage depth in naive
 K-means clustering of promoter H3K27me3 relative coverage depth in naive
  day 0 samples.
  day 0 samples.