Fig. 5: TRIM25 negatively regulates tumor cells’ ROS and apoptosis in cooperation with Nrf2.

a Western blot analysis of the levels of cleaved caspase3 in HCT116 and Huh7 cells for control (shNC) or after stable knockdown of TRIM25. b, e Western blot analysis of the levels of cleaved caspase3 in Huh7 cells for stalely knockdown (b) or overexpression (e) of TRIM25 treated with TM (5 μg/ml) or TG (1 μM) for 12 h. c, g Quantification of apoptotic HCT116 cells stable knockdown (c) or overexpression (g) of TRIM25, treated with or without TG (1 μM) for 12 hand analyzed by flow cytometry. f Western blot analysis of the levels of cleaved caspase3 in control and F-TRIM25-expressing HCT116 cells, treated with or without TG (1 μM) for 12 h. d, h Quantification of apoptotic Huh7 cells stable knockdown (d) or overexpression (h) of TRIM25, treated with TM (5 μg/ml) or TG (1 μM) for 12 h and analyzed by flow cytometry. i Western blot analysis of the efficiency of the stable cell knocks down of Nrf2 in HCT116 and Huh7 cells. j Quantification of ROS level and apoptotic HCT116 cells expressing control or F-TRIM25 and with simultaneously knockdown of Nrf2, treated with or without TG (1 μM) for 12 h. k, m Quantification of ROS level (k) and apoptotic (m) Huh7 cells expressing control or F-TRIM25 and with simultaneously knockdown of Nrf2, treated with TM (5 μg/ml) or TG (1 μM) for 12 h. l Western blot analysis of the levels of cleaved caspase3 in control and F-TRIM25-expressing HCT116 cells after stable knockdown of Nrf2, treated with or without TG (1 μM) for 12 h. For c, d, g, h, j, k, and m, data represent the mean ± SEM (n = 3). Statistical significance was assessed using two-tailed Student’s t-tests. n.s. not significant.