Figure 7.

G6PD downregulation interferes with the transformed phenotype of ERMS cells but its overexpression does not affect miR-206-mediated ERMS cells differentiation. (A) Western Blot analysis of G6PD in RD18 cells conditionally expressing a control (shctrl) or a G6PD-directed (shG6PD) shRNA, treated or not with doxycycline for the indicated days (shRNA not induced, NI; shRNA induced, IND). (B) MTT analysis of the cells indicated in (A). Cells were analyzed for the indicated days in absence of doxycycline (shRNA not induced, NI) or after doxycycline administration (shRNA induced, IND). The number of cells at day 0 was set at 100%. (C) Quantification of soft agar growth assays of the cells indicated in (A). The number of colonies obtained from cells maintained in absence of doxycycline (shRNA not induced, NI) was set at 100%. (D) Representative images of the experiment indicated in (C). (E) MTT analysis of RD18 NpBI-206 cells overexpressing or not G6PD lacking the 3’UTR. Cells were analyzed for the indicated days in the absence of doxycycline (miR-206 not induced, NI) or after doxycycline administration (miR-206 induced, IND). The number of cells at day 0 was set at 100%. Student's t-test was used to evaluate statistical significance: *P <0.05. (F) Western blot analysis of the indicated proteins in the cells indicated in (E), treated or not with doxycycline for the indicated days (miR-206 not induced, NI; miR-206 induced, IND).