Research Paper Volume 16, Issue 20 pp 12977—13011

Figure 5. H3K9me3 heterochromatic mark is reduced on satellite tandem repeats in WRN-depleted cells. (A) Quantitation of IF in situ for H3K9me3 levels in WI38hTERT expressing shRNA against WRN or non-specific control (NS). Crossbars are distribution means. The graph represents three independent experiments. (B) ChIP qPCR analyses of H3K9me3 levels on the indicated genomic loci, performed in the same cells as in (A). The graph summarizes results of five independent experiments with technical triplicates. (C) ChIP qPCR analyses of histone H3 levels on the indicated genomic loci, performed in the same cells as in (A). The graph summarizes results of three independent experiments with technical triplicates. In (B-C), red triangles are means, and boxplots mark first and fourth quartiles and the distributions’ medians. (D) Quantitations of the indicated PLA in situ analyses in WI38hTERT cells expressing shRNA against WRN or control shRNA. Graphs represent two (HP1α/KAP1) and four (HP1α/LBR) independent experiments each. (E) Quantitations of the indicated IF in situ analyses performed on the same cells as in (D). Graphs represent two (KAP1), and three (HP1α) independent experiments each.