File S1 - Inactivating UBE2M Impacts the DNA Damage Response and Genome Integrity Involving Multiple Cullin Ligases

Containing the following supporting information files: Figure S1. Comet assay. The tail moment is the length of the tail times the density of the tail. % tail DNA is the density of the tail divided by the density of the tail plus the density of the head. Figure S2. Growth suppression by UBE2M silencing is enhanced by PARP inhibitor ABT-888 in HEY cells. Figure S3. HR repair was was performed upon knockdown of UBE2M (two siRNAs) and CHK1. Figure S4. Expression level of GFP is unaffected by UBE2M knockdown. HeLa cells that stably express pLKO vector or pLKO-UBE2M shRNA were transiently transfected with pEGFP-N1 vector. 48 hours after, cells were harvested for flow cytometer analysis. Figure S5. MLN4924 or knockdown of IkB does not significantly affect the level of RAD51 or FANCD2 proteins. HeLa cells were simultaneously treated with siRNAs against IkB-α and IkB-β, then treated with MLN4924, for western blot analysis. Figure S6. Knockdown of UBE2M inhibits neddylation of Cullins. Figure S7. Knockdown of CUL2 arrests the cell cycle at the G1-S boundary. Left: confirmation of two independent siRNAs. Right: Double thymidine experiments were performed using the two siRNAs against CUL2 in HEY cells. Figure S8. Confirmation of knockdown for indicated siRNAs. (EPS)