Fig. 2: GLIS2 negatively regulates the expression of PUMA.
From: GLIS2 promotes colorectal cancer through repressing enhancer activation
a HCT116 cells were transfected with two different siRNAs of GLIS2 for 48 h followed by treatment with 375 μM 5-FU for 8 h, and the expression of PUMA, p21, and TP53 was determined by qRT-PCR. Data are presented as mean ± SEM of three independent experiments. b GLIS2 was knocked down with two different siRNAs in HCT116 and cell lysates were immunoblotted with indicated antibodies. c, d GLIS2 was exogenous expressed in HCT116 and the expression of PUMA was determined by qRT-PCR (c) and western blotting (d). PUMA protein abundance was quantified by ImageJ. e DEGs between control and GLIS2 knockdown under DMSO or 5-Fu conditions were combined as siGLIS2 DEGs. Venn diagram shows the overlapped gene number between siGLIS2 DEGs and p53 target genes. f Heatmap of the overlapped genes in (e). Genes validated with RT-PCR were in red. g HCT116 p53 knockout cells were prepared as in 2 A and the expression of PUMA was determined by qRT-PCR. *p-value ≤ 0.05, **p-value ≤ 0.01 (t-test). Histograms are presented as mean ± s.d. of three biological replicates.
