Gene expression changes after depletion of Cyclin F and atypical E2Fs in HeLa cells.

Using RNAi technology, Cyclin F, E2F7+8, or the combination of all proteins in were knocked down HeLa cells. The goal was to study which gene expression programmes are altered by cyclin F depletion in G2/M cells. Furthermore, we had performed experiments suggesting that cyclin F mediates degradation of the atypical E2F repressor proteins E2F7 and E2F8. Therefore we investigated using combinatorial knockdown if additional depletion of E2F7/8 rescues gene expression changes caused by cyclin F. Our resuilts show that cyclin F depletion downregulates expression hundreds of E2F target genes, which could be prevented by additional E2F7/8 depletion. Cells were harvested 48 hours after transfection. Final concentration of each individual siRNA was 10 nM. Nocodazole (250 ng/mL) was added during the last 16 hours to collect all cells in G2/M-phase. Per time point we used 4 biological replicates. Control cells received a scrambled siRNA.