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. 2011 Jul 8;39(19):8329–8341. doi: 10.1093/nar/gkr529

Figure 2.

Figure 2.

Requirements for H3.3 and HP1γ in HSP70 transcription. (A) HeLa cells were stably transfected with shRNAs targeting to non-specific control, H3.3 or HP1γ. The efficiency and selectivity of knockdown were determined under non-heat shock and heat shock conditions by western blotting (upper panel) and qRT–PCR (lower panel). All transcription levels were normalized to that of GAPDH. Note that H3.3 protein is encoded by two different genes, H3.3 A and H3.3B. β-Actin was used as an internal control for equal loading in all western blot analyses. (B) Whole-cell extracts were prepared for analyses of HSP70 expression in these cells by western blotting analysis. Total mRNA was purified and subjected to qRT–PCR to examine transcription levels of HSPA1 and HSPA6 genes under non-heat shock and heat shock conditions. Average and SD of three independent experiments are shown.