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Fig. 4 | Clinical Epigenetics

Fig. 4

From: Regional gain and global loss of 5-hydroxymethylcytosine coexist in genitourinary cancers and regulate different oncogenic pathways

Fig. 4

Increased stemness and DNA 5hmC in 3D-cultured prostate cancer cells. A Formation and growth of spheroids by 3D-cultivating 22RV1 cells for 7 consecutive days; scale bar = 10 µm. B, C Quantification of the stem cell marker expression in 22RV1 and PC3 cell lines by real-time PCR. Cells cultured in 2D rigid dishes were used as control. D–F Subcutaneous xenografts (D) derived from 2D- to 3D-cultured 22RV1 cells, the growth kinetics (E) determined by measuring tumor volume and corresponding tumor weights (F). Each group n = 6. G, H Dot blot (G) and hMeDIP-seq (H) showing the 5hmC alteration introduced by the 3D culture in prostate carcinoma cell lines 22RV1 and PC3. I 5hmC density at hyper-DhMRs shared by 3D-cultured 22RV1 (left) and PC3 (right) cells. J Functional relevance of the hyper-hydroxymethylation introduced by 3D-culturing of 22RV1 (left) and PC3 (right) cells. K Genome browser tracks depicting the 5hmC surge of MYC attributed to 3D-culturing. In (B), (C), (E) and (F), 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.

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