Fig. 6

Inhibition of the growth of tumor stem cell-like cells by vitamin C. A Dot blot of 5hmC levels in prostate carcinoma cell lines 22RV1 and PC3 receiving APM treatment and vehicle-treated control cells. B 5hmC density at hyper-DhMRs shared by APM-treated 22RV1 (left) and PC3 (right) cells. C Proliferation curves of 22RV1 and PC3 cells in medium supplemented with increasing concentrations of APM. D, E 22RV1 and PC3 spheroids cultivated in 3D soft gel with the presence of increasing concentrations of APM at day 0 and day 5 (E) and the growth kinetics over 7 consecutive days (F); scale bar = 10 µm. F Bar plots showing the influence of APM treatment for 7 days on the number of 22RV1 and PC3 spheroids. G Heatmap of Pearson correlation values that were generated on the 5hmC patterns at 100-bp genomic windows of the indicated samples. H Genome browser tracks depicting 5hmC levels of CARD14 in normal prostate samples, prostate cancer cells, prostate cancer cells treated with APM and prostate cancer cells cultured at 3D systems. I, J Bar plots displaying 5hmC (I) and gene expression levels (J) of CARD14 in APM- and vehicle-treated 22RV1 and PC3 cells cultured in 2D and 3D systems. In (C), (F), (G), (I) and (J), error bars represent mean ± standard deviation. P values were produced with t test. ****P < 0.0001; ***P < 0.001; **P < 0.01;*P < 0.05