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. 2020 May 21;10:667. doi: 10.3389/fonc.2020.00667

Figure 3.

Figure 3

Ectopic expression of ten-eleven translocation 1 (TET1)-CD and vitamin C treatment reduce urinary bladder cancer (UBC) cell proliferation and invasion, dependent on its catalytic domain. (A) Confirmation of overexpression of TET1-CD by Western blotting. (B) Dot blot assay of 5-hydroxymethylcytosine (5 hmC) level in TET1-CD and TET1-CDmut overexpression T24 cells, as well as empty vector (EV) control cells with DNA concentration gradients. (C–F) Effects of TET1 overexpression on cell viability (C), colony formation (D), migration (E), and invasion (F) in vitro. (G) Detection of 5 hmC levels of T24 cell treated with various doses of vitamin C by dot blot assay. (H) 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay of T24, 5,637 UBC and nonmalignant SV-HUC-1 urothelial cell viabilities at various concentrations with vitamin C treatment for 48 h. (I) Colony formation assay for T24 and 5,637 cells at various concentrations of vitamin C. (J) Apoptosis assay of T24 cells treated with various concentrations with vitamin C for 48 h. Scale bar, 200 μm (E) and 100 μm (F). **p < 0.01, ***p < 0.001.