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Figure 6: Effects of Stampidine on Methylation Levels of HIV-Responsive Transcription Factor Genes. HIV-Induced Dynamic Increases in Expression of Methylated Transcription Factor Genes. A total of 20 genes showing dynamic increases in expression after HIV infection were methylated by Stampidine. Gene expression values (signal difference metric) were mean centered to 24 h time point post-HIV exposure in the presence and absence of Stampidine to assess genes dynamically induced in expression by the virus at 48 h. One-way agglomerative hierarchical clustering technique was used to organize gene expression patterns across treatments to identify genes with similar profiles. The heat map depicts genes upregulated (red) vs. downregulated (green) at 48 h relative to 24 h (A). Gene expression values for GNAS were scales by a factor of 0.2 to a comparable dynamic range observed with other gene transcripts. Stampidine prevented HIV-induced upregulation of 19 of these 20 genes. The effects of Stampidine on gene methylation levels was determined in 3 T-cell lines (LOUCY, CEM, MOLT3) by comparing the methylation levels in samples treated with Stampidine to those in vehicle-treated control samples. Methylation profile depicts the increase in methylation in the presence of Stampidine for methylation probes corresponding to these 20 genes (B). A total of 20 genes showing dynamic increases in expression after HIV infection were methylated by Stampidine. See Table S7 for details.
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