Fig. 4: Ectopic expression of GLI1 reverses the phenotype caused by UHRF1 silencing.

A Gene Ontology analysis of downregulated genes in the CRL-8024 UHRF1 knockdown group compared with the CRL-8024 control group. B Heatmap displays stemness- and differentiation-related genes that were differentially expressed between CRL-8024 control and UHRF1 knockdown cells. The color of each cell shows the Z score (log2 of relative abundance scaled by SD) of the mRNA in that sample. C Venn diagram among three datasets in CRL-8024 cells. D WB analysis of UHRF1, GLI1, CD44, and CD133 protein expression in CRL-8024 cells with UHRF1 knockdown (shUHRF1-EV) and subsequent GLI1 ectopic expression (shUHRF1-GLI1). EV: empty vector. E CCK8 assay was used to assess the viability of the indicated stable CRL-8024 cell lines. F Representative images and quantification of foci formation induced by the indicated CRL-8024 cells (n = 3). G Representative images and statistical results of the transwell assay of the indicated CRL-8024 cells (n = 3). H Representative images and quantification of spheroids formed by the indicated stable CRL-8024 cell lines (n = 3). I The CSC subpopulations were evaluated by flow cytometry. CRL-8024 cells were stained with anti-CD133-PE and anti-CD44-APC antibodies. The percentages were calculated and depicted in the bar chart (n = 3). Mean ± SD. P values were determined using unpaired Student’s t test. ^∗P < 0.05, ^∗∗P < 0.01, ^∗∗∗P < 0.001. Ns not significant.