Figure 3.

HCQ inhibited HCC cancer stem-like cells’ self-renewal properties. (A) Correlation analysis of TLR9 expression with stem cell marker expression. (B) HCQ inhibited the tumor sphere-forming capacity (scale bar 200 μm) and (C) colony-forming capacity in HCC (Huh7 and HepG2) cells. The bar plot represents the quantification of tumorsphere and colony-forming abilities after HCQ treatment calculated by ImageJ, provided in (B) (below) and (C) (below), respectively. (D) Western blot analysis expression of TLR9 and cancer stem/self-renewal-related marker (Oct4) or GAPDH (loading control). Representative protein bands are shown a reduction in expression TLR9 and Oct4 observed after the HCQ treatment (10 μM). (E) mRNA expression of TLR9, CD44, Oct44, and KLF4 was measured by qRT-PCR; GAPDH was used to normalize the expression levels. Data are the mean ± SEM of three independent experiments. * p < 0.05.