Figure 4. Hedgehog and Notch signaling were involved in ALDH^hiCD44^hi TIC maintenance.

A and B, mRNA levels of Hedgehog and Notch signaling genes by QPCR in sorted cells from H1650 and HCC827. C, mRNA levels of HES1 or GLI1 by QPCR in HCC827 after Notch inhibition by RO4929097 or Hedgehog inhibition by cyclopamine. D and E, Proportions of ALDH⁺CD44⁺ by flow cytometry in HCC827 and H1650 after pathway inhibition for 1 or 4 days. F, Spheroid formation assay of HCC827 cells treated with inhibitors. *, p < 0.05; **, p < 0.01; ***, p < 0.001, compared with unsorted or control; #, p < 0.05; ##, p < 0.01, compared with ALDH^hiCD44^hi. Data represent mean ± SD of triplicate experiments.