Fig. 4: Investigation and confirmation of target molecule alterations by a cDNA microarray in ATO-treated HCC cells.

a The microarray heat map (top) shows some of the profiles of differentially expressed mRNAs in two cells with over two-fold changes. Pseudo-colors indicate differential expression (green, transcript levels below the control; red, transcript levels greater than the control). Bottom: confirmation of some of the major candidate mRNAs by qRT-PCR analysis. *P < 0.05, **P < 0.01, determined by two-sided Student’s t-test. b GO analysis of differential expressing mRNAs classified by their biological process for MCM7 in HCC cells after ATO treatment. c qRT-PCR analysis of expressions of MCM2, MCM3, MCM4, MCM5, and MCM6 in two cells after ATO treatment. *P < 0.05, **P < 0.01, determined by two-sided Student’s t-test. d Western blot (WB) analysis of MCM7 expression in HCC cell lines for 48 h of treatment with 3.6 μM ATO (top). Bottom shows MCM7 expression in HCC cell lines treated with ATO at the indicated dosages. e qRT-PCR analysis of MCM7 expression levels in tumorspheres (n = 3) treated with ATO. WT, wild type adherent cells; Spheres-con, tumorspheres without ATO treatment; Spheres-ATO, tumorspheres with ATO treatment. *P < 0.05, **P < 0.01, determined by two-sided Student’s t-test. f H&E staining and MCM7 IHC staining of tumors derived from the mice that received local injection of ATO or PBS. Bars, 100 μm. **P < 0.01, *P < 0.05, determined by two-sided Student’s t-test