Figure 3: The effect of HDAC1 and HDAC2 inhibition depends on Gli K518 acetylation. | Scientific Reports

Figure 3: The effect of HDAC1 and HDAC2 inhibition depends on Gli K518 acetylation.

From: Selective targeting of HDAC1/2 elicits anticancer effects through Gli1 acetylation in preclinical models of SHH Medulloblastoma

Figure 3

(a) QPCR analysis of Gli1 mRNA levels in MEF Sufu−/− and MEF Ptch1−/− treated with 0.5 μM MGCD0103 for 24 hours. Each experiment was performed in triplicate. n = 3 (b) WT or Gli1 and Gli2 KO MEF cells were treated with SAG for 48 h and/or 0.5 μM MGCD0103 for the last 16 h and QPCR analysis of Ptch1 mRNA levels was performed in triplicate. n = 3 (c) QPCR analysis of Ptch1 mRNA in Med1-MB cells expressing siRNA for Gli1 (siGli1) or si control RNA (siCTR) for 96 h and then treated with 0.5 μM MGCD0103 for 24 h. Upper panel, western blot analysis indicating the efficacy of Gli1 knockdown. Vinculin was used as loading control. n = 4 (d) Growth curve of Med1-MB cells expressing siRNAs targeting Gli1 (siGli1) or control siRNA (siCTR) for 96 h and then treated with 0.5 μM MGCD0103 or DMSO for the indicated time. Each experimental point was performed in triplicate. n = 3 (e) Western blot analysis of p21 in Med1-MB cells expressing siGli1 or siCTR RNA for 96 h and treated with 0.5 μM MGCD0103 or DMSO as a control for 48 h. Gli1 levels are shown as control of knockdown efficacy. (f) Acetylation of endogenous Gli1 in Med1-MB treated with 0.5 μM MGCD0103 or 100 nM vorinostat or DMSO. Total Gli1 levels are shown (g) Growth curve in DAOY cells stably transfected with Gli1 WT or Gli1 K518R mutant. Cells were seeded in triplicates, treated with MGCD0103 or DMSO as control and counted at the indicated time points. n = 3 *p < 0.05; **p < 0.01. Uncropped Western blot gels related to this figure are displayed in Suppl. Fig. 6.

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