Fig. 5: mtROS clearance attenuates the oncogenic function of SQLE.

A Intracellular ROS levels as assessed by flow cytometry in UMUC3 and J82 cells with or without SQLE overexpression (n = 3, performed in triplicate). The impact of Mito-TEMPO (TEMPO) (40 µM) treatment on the viability of UMUC3 and J82 cells with SQLE overexpression, as determined by growth curves (B) (n = 3, performed in triplicate), colony formation assays (C) (n = 3, performed in triplicate), wound healing assays (D) (n = 3, performed in triplicate), and transwell assays (E) (n = 3, performed in triplicate). F A schematic diagram for in vivo Mito-TEMPO treatment. G Representative gross morphology and H&E staining of the bladder following in vivo Mito-TEMPO treatment (left). The effects of Mito-TEMPO on BCa formation are illustrated by tumor incidence and disease severity (right). (Rosa26-Sqle mice treated with NS controls (n = 5), Sqle bladTg micetreated with NS controls (n = 4), Sqle bladTg mice treated with Mito-TEMPO (n = 4)). H Ki-67 staining of the bladder from Rosa26-Sqle mice treated with NS controls (n = 5), Sqle bladTg micetreated with NS controls (n = 4), and Sqle bladTg mice treated with Mito-TEMPO (n = 4). I Western blots analysis of PCNA expression in the bladder from Rosa26-Sqle mice treated with NS controls, Sqle bladTg mice treated with NS controls, and Sqle bladTg mice treated with Mito-TEMPO. Data are represented as means ± SD. *P < 0.05, **P < 0.01, ***P < 0.001. Unpaired, two-tailed Student’s t-test (A, C, D, E, H, I). Difference in cell viability between the two groups was evaluated through ANOVA with repeated-measures analysis of variances (B). Scale bars, 500μm (low magnification) and 200 μm (higher magnification) (G). Scale bars, 200 μm (H).