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. 2020 Feb 17;9(2):23. doi: 10.1038/s41389-020-0202-7

Fig. 5. Effect of ZIC2 and GLI1 siRNA knock down on viability, survival, and stem cell markers expression in CTBPH1 cells.

Fig. 5

a Spheroid formation assay for CTBPH1 cells transfected with scrambled (scr) and ZIC2 siRNA. b CTBPH1 cells transfected with scr or ZIC2 siRNA were incubated with or without Cd for 24 h. Cell lysates were subjected to western blotting using antibodies against ZIC2, GLI1, SOX2, CD44, p65, Notch1, and β-actin. c RNA isolated from CTBPH1 cells transfected with scr or ZIC2 siRNA and incubated with or without Cd for 24 h was subjected to qRT-PCR analysis using ZIC2, SOX2, Nanog, CD44, and Notch1 primers. d Cell viability of CTBPH1 cells treated with scr or ZIC2 siRNA was determined by trypan blue assay. e ZIC2, GLI1, p65, and cleaved PARP expression in cell lysates prepared from scr and ZIC2 siRNA-treated CTBPH1 cells. f Cell viability of CTBPH1 treated with scr or GLI1 siRNA: The cell viability was determined by trypan blue assay. g Cell lysates were prepared from scr and GLI1 siRNA-treated CTBPH1 cells to determine the protein levels of ZIC2, GLI1, p65, and cleaved PARP by western blot analysis. #, not significant, *p < 0.05, **p < 0.01, and ***p < 0.001; *, nonspecific band.