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. 2018 Jan 24;24:461–472. doi: 10.12659/MSM.908430

Figure 2.

Figure 2

DNMT and HDAC inhibition decrease CSLCs self-renewal. (A) Western blot was used to analyze DNMT1/3b and HDAC1/3 protein levels following 5-AzaC or TSA treatment (left). Respective changes in DNMTs (middle) and HDACs (right) protein levels. (B) RT-PCR analysis of DNMT1 and DNMT3b mRNA levels with 5-AzaC treatment. (C) Total HDAC activity of CSLCs with TSA treatment. (D) RT-PCR analysis of HDAC1 and HDAC3 mRNA levels with TSA treatment. (E) Representative images of spheres (10×, left) and sphere counts (right) in first- and second-generation in cells with 5-AzaC or TSA treatment. (F) RT-PCR analysis of stemness-associated genes in sphere cells with 5-AzaC or TSA treatment (groups of 5-AzaC, TSA and 5-AzaC+TSA vs. DMSO group). Results were normalized to expression of GAPDH. (G) Flow cytometry analysis of CD133 cell surface expression in sphere cells with 5-AzaC or TSA treatment. Data represent mean ±SD from at least 3 independent experiments, * p<0.05; ** p<0.01; ***p<0.001.