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@@ -98,31 +98,6 @@ mechanisms, or a diagram for periodic checking. -->
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:::
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-## My thesis topics
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-
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-<!-- TODO: Needs revision -->
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-
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-### Topic 1: Immune memory
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-Genome-wide epigenetic analysis of H3K4 and H3K27 methylation in naïve
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-and memory $\mathsf{CD4}^{+}$ T-cell activation
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-
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-### Topic 2: Diagnostics for rejection
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-Improving array-based diagnostics for transplant rejection by
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-optimizing data preprocessing
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-
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-### Topic 3: Blood profiling during treatment
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-Globin-blocking for more effective blood RNA-seq analysis in primate
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-animal model for experimental graft rejection treatment
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-
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-## Today's focus
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-
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-### \Large Topic 1: Immune memory
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-
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-\Large
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-
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-Genome-wide epigenetic analysis of H3K4 and H3K27 methylation in naïve
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-and memory $\mathsf{CD4}^{+}$ T-cell activation
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-
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## Memory cells: faster, stronger, and more independent
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@@ -152,8 +127,6 @@ Compared to naïve cells, memory cells:
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Result:
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-\normalsize
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-
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* Memory cells require progressively higher doses of immune suppresive
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drugs
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* Dosage cannot be increased indefinitely without compromising the
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@@ -161,12 +134,38 @@ Result:
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:::
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+## 3 problems relating to transplant rejection
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+
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+### 1. How are memory cells different from naïve?
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+
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+\onslide<2->{Genome-wide epigenetic analysis of H3K4 and H3K27 methylation in naïve
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+and memory $\mathsf{CD4}^{+}$ T-cell activation}
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+
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+### 2. How can we diagnose rejection noninvasively?
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+
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+\onslide<3->{Improving array-based diagnostics for transplant rejection by
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+optimizing data preprocessing}
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+
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+### 3. How can we evaluate effects of a rejection treatment?
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+
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+\onslide<4->{Globin-blocking for more effective blood RNA-seq analysis in primate
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+animal model for experimental graft rejection treatment}
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+
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+## Today's focus
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+
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+### \Large 1. How are memory cells different from naïve?
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+
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+\Large
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+
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+Genome-wide epigenetic analysis of H3K4 and H3K27 methylation in naïve
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+and memory $\mathsf{CD4}^{+}$ T-cell activation
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+
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## We need a better understanding of immune memory
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* Cell surface markers fairly well-characterized
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* But internal mechanisms poorly understood
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-
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+
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. . .
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\vfill
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@@ -177,51 +176,66 @@ Result:
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histone modification is involved in $\mathsf{CD4}^{+}$ T-cell
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activation and memory.
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-## Experimental design
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+## Which histone marks are we looking at?
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-* Separately isolate naïve and memory $\mathsf{CD4}^{+}$ T-cells from
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- 4 donors
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-* Activate with CD3/CD28 beads
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-* Take samples at 4 time points: Day 0 (pre-activation), Day 1 (early
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- activation), Day 5 (peak activation), and Day 14 (post-activation)
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-* Do RNA-seq + ChIP-seq for 3 histone marks (H3K4me2, H3K4me3, &
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- H3K27me3) for each sample.
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+. . .
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-Data generated by Sarah Lamere, published in GEO as
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-[GSE73214](https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE73214)
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+::: incremental
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-## Time points capture phases of immune response
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+* **H3K4me3:** "activating" mark associated with active transcription
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+
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+* **H3K4me2:** Correlated with H3K4me3, hypothesized "poised" state
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+
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+* **H3K27me3:** "repressive" mark associated with inactive genes
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+
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+:::
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+
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+. . .
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+
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+\vfill
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+
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+All involved in T-cell differentiation, but activation dynamics
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+unexplored
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+
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+## ChIP-seq measures DNA bound to marked histones[^chipseq]
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\centering
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-<!-- { height=75% } -->
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+{ height=70% }
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+
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+[^chipseq]: [Furey. "ChIP-seq and beyond: New and improved methodologies to detect and characterize protein-DNA interactions". In: Nature Reviews Genetics (2012)](http://www.nature.com/articles/nrg3306)
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-## Why study these histone marks?
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+## Experimental design
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::: incremental
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-* **H3K4me3:** "activating" mark associated with active transcription
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+* Separately isolate naïve and memory $\mathsf{CD4}^{+}$ T-cells from
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+ 4 donors
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-* **H3K4me2:** Correlated with H3K4me3, hypothesized as a "poised" state
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+* Activate with CD3/CD28 beads
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-* **H3K27me3:** "repressive" mark associated with inactive
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+* Sample at 4 time points: Day 0 (pre-activation), Day 1 (early
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+ activation), Day 5 (peak activation), and Day 14 (post-activation)
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-* All 3 involved in T-cell differentiation, but activation dynamics
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- unexplored
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+* RNA-seq + ChIP-seq of 3 histone marks (H3K4me2, H3K4me3, & H3K27me3)
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+ for each sample.
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:::
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-## ChIP-seq sequences DNA bound to marked histones[^chipseq]
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+Data generated by Sarah Lamere, published in GEO as
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+[GSE73214](https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE73214)
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-\centering
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+## Time points capture phases of immune response
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-{ height=70% }
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+\centering
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-[^chipseq]: [Furey. "ChIP-seq and beyond: New and improved methodologies to detect and characterize protein-DNA interactions". In: Nature Reviews Genetics (2012)](http://www.nature.com/articles/nrg3306)
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+
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## A few intermediate analysis steps are required
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-
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+\centering
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+
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+
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## Histone modifications occur on consecutive histones
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@@ -231,15 +245,13 @@ Data generated by Sarah Lamere, published in GEO as
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## Histone modifications occur on consecutive histones
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-
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-
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-## Histone modifications occur on consecutive histones
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-
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-
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-
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-## Histone modifications occur on consecutive histones
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-
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-
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+\begin{figure}
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+\centering
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+\only<1>{\includegraphics[width=\textwidth,height=0.7\textheight]{graphics/presentation/CCF-plots-A-SVG.png}}
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+\only<2>{\includegraphics[width=\textwidth,height=0.7\textheight]{graphics/presentation/CCF-plots-B-SVG.png}}
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+\only<3>{\includegraphics[width=\textwidth,height=0.7\textheight]{graphics/presentation/CCF-plots-C-SVG.png}}
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+\caption{Strand cross-correlation plots show histone-sized wave pattern}
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+\end{figure}
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## SICER identifies enriched regions across the genome
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@@ -263,23 +275,15 @@ Data generated by Sarah Lamere, published in GEO as
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## Peaks in promoters correlate with gene expression
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-
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-
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-## Peaks in promoters correlate with gene expression
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-
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-
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-
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-## Peaks in promoters correlate with gene expression
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-
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-
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-
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-## Peaks in promoters correlate with gene expression
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-
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-
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-
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-## Peaks in promoters correlate with gene expression
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-
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-
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+\begin{figure}
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+\centering
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+\only<1>{\includegraphics[width=\textwidth,height=0.7\textheight]{graphics/presentation/FPKM-by-Peak-Violin-Plots-A-SVG.png}}
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+\only<2>{\includegraphics[width=\textwidth,height=0.7\textheight]{graphics/presentation/FPKM-by-Peak-Violin-Plots-B-SVG.png}}
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+\only<3>{\includegraphics[width=\textwidth,height=0.7\textheight]{graphics/presentation/FPKM-by-Peak-Violin-Plots-C-SVG.png}}
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+\only<4>{\includegraphics[width=\textwidth,height=0.7\textheight]{graphics/presentation/FPKM-by-Peak-Violin-Plots-D-SVG.png}}
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+\only<5>{\includegraphics[width=\textwidth,height=0.7\textheight]{graphics/presentation/FPKM-by-Peak-Violin-Plots-Z-SVG.png}}
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+\caption{Expression distributions of genes with and without promoter peaks}
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+\end{figure}
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## The story so far
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