Fig. 4

Effects of TIC-related signaling inhibitors on the cell viability of HCC PDOs. a Sorafenib treatment affected the CD44 levels at the protein or RNA levels in HCC cell lines; b 4 HCC PDOs were treated with a Notch signaling inhibitor (YO-01027), Wnt signaling inhibitor (Wnt-C59), Hippo signaling inhibitor (verteporfin), or Hedgehog signaling inhibitor (GANT61) for 6 days and cell viability was recorded. c Immunofluorescence images of HCC PDOs after treatment with DMSO or GANT61 for 6 days. The cells were stained for TUNEL (green) and stained with DAPI (blue). Scale bar: 50 μm. d Immunofluorescence images of HCC PDOs after treatment with DMSO or GANT61 for 6 days. The cells were stained for Ki-67 (green) and stained with DAPI (blue). Scale bar: 50 μm. e Protein expression of Oct4, Sox2 and Nanog in HCC PDOs after treatment with GANT61 (10 μM) determined by Western blotting. (** p < 0.01, *** p < 0.001, P < 0.05 is considered statistically significant)