Fig. 3

The overexpression of Notch1 reverses the FXR agonist-induced asymmetric cell division in liver CSCs. a Huh7 cells were transfected with Notch1 expression plasmid or vector plasmid, and quantitative real-time PCR confirmed Notch1 overexpression. b Huh7 cells were transfected with Notch1 expression plasmid or vector plasmid, Western blottings confirmed Notch1 overexpression, normalized to GAPDH. c After transfected with Notch1 expression plasmid or vector plasmid for 7 h, Huh7 cells were treated with GW4064 (10 μM) for another 24 h, and quantitative real-time PCR was assayed for SHP and Notch1 expression. d After transfected with Notch1 expression plasmid or vector plasmid for 6 h, Huh7 cells were treated with GW4064 (10 μM) for another 36 h, and Western blotting was assayed for Notch1 and NICD1 expression, normalized to LaminB or GAPDH. e Left: Representative images of the paired-cell assay of liver CSCs from Huh7 cells. BrdU, red; DNA, blue; Sox9, green. Scale bars: 5 μm. Right: Quantification of BrdU asymmetry or symmetry in liver CSCs after Notch1 overexpression expression maintained treated with GW4064 (10 μM) medium. Data were means of three separated experiments ± SEM, *p < 0.05, **p < 0.01