TMZ-induced autophagy is MMR and HR dependent.

<p>(<b>a</b>) Autophagy induction after TMZ treatment determined by MDC staining and quantified by flow cytometry in LN-229 cells transiently knocked-down for MSH6 (or transfected with scrambled siRNA). Insert: MSH6 expression in LN-229 cells transiently transfected with MSH6 or scrambled siRNA 96 h after transfection performed by western blot, ERK-2 was used as loading control. (<b>b</b>) MDC positive cells determined 96 h after TMZ treatment (100 µM) via flow cytometry in stable MSH6 expressing cells (DLD-1-MSH6) compared to control cells. Insert: MSH6 expression in overexpressing clone and control performed by western blot, ERK-2 was used as loading control. (<b>c</b>) MDC positive cells in HCT-116 and HT-29 cells, as determined 96 h after TMZ treatment. (<b>d</b>) Rad51 expression in stable Rad51 knockdown glioma cells (LN-229 Rad51shRNA) compared to empty vector transfected cells (LN-229-pS-empty) performed by western blot, ERK-2 was used as loading control. (<b>e</b>) Rad51 foci (yellow) 72 h after TMZ treatment (20 µM) in knockdown clone and control. TO-PRO-3 was used for nuclear staining (<b>f</b>) MDC positive cells determined 96 h after TMZ treatment (10 µM) via flow cytometry in knockdown clone and control.