Figure 2.

SESN2 was required for autophagic induction and anchorage-independent growth inhibition by ISO in human bladder cancer UMUC3 cells. (A) UMUC3 cells were treated with various concentrations of ISO for 24 h. The cells were extracted and cell lysates were subjected to western blotting to detect the expression of BECN1, SESN2 and LC3. (B and D) shRNA BECN1 (B) and shRNA SESN2 (D) were stably transfected into UMUC3 cells, and the stable transfectants were identified. (C and E) The indicated stable transfectants were subjected to ISO treatment for 24 h for determination of LC3 in ISO-induced autophagy in UMUC3 cells. (F) Representative images of colonies of UMUC3 (shSESN2) and UMUC3 (Nonsense) cells in soft-agar assay in the presence or absence of various concentrations of ISO were captured using a microscope. (G) Colonies shown in (F) were counted under a microscope with more than 32 cells of each colony. The results are presented as colonies per 10⁴ cells, and the bars shows mean ± SD from 3 independent experiments. The symbol (*) indicates a significant decrease from vehicle control (p < 0.05).