Fig. 4

FXR promotes asymmetric division of liver CSCs through regulation of Notch1. a The expression of FXR was examined by quantitative real-time PCR and Western blotting, normalized to GAPDH. b After interfering FXR expression for 12 h, Huh7 cells were treated with GW4064 (10 μM) or control DMSO for another 24 h, and quantitative real-time PCR was assayed for SHP and Notch1 expression. c After interfering FXR expression for 12 h, Huh7 cells were treated with GW4064 (10 μM) or Control DMSO for another 36 h, and Western blotting was assayed for Notch1 and NICD1 expression, normalized to LaminB or GAPDH. d Left: Representative images of the paired-cell assay of liver CSCs from Huh7 cells. BrdU, red; DNA, blue; CD133, green. Scale bars: 5 μm. Right: Quantification of BrdU asymmetry or symmetry in liver CSCs after interfering FXR expression maintained treated with GW4064 (10 μM) medium. e Left: Representative images of the paired-cell assay of liver CSCs from Huh7 cells. BrdU, red; DNA, blue; Notch1, Green. Scale bars: 5 μm. Right: The percentage of BrdU and Notch1 co-expression or inverse expression in daughter cells of CD133⁺ liver CSCs from Huh7 cells undergoing division. Data were means of three separated experiments ± SEM, *p < 0.05, **p < 0.01, ***p < 0.001