siRNA Transfection by Electroporation

Small interference (si)RNAs were purchased from Eurofins MWG. Transfection was performed by electroporation as described [66 (link)]. Briefly, cells were trypsinized, washed once in RPMI medium, electroporated by using a Bio-Rad GenePulserX-Cell at 250 V and 800 μF and kept on ice for 30 min. Cells were then transferred to a 6 or 10 cm (Ø) culture dish and incubated for two days to allow the siRNA to downregulate the expression of its target. The effect of the siRNA was confirmed by Western blot and/or Q-RT-PCR analysis. The following siRNAs (sense-strand) were used: siBcl3 (5′-UGG UCU UCU CUC CGC AUC A-3′), siLuc (5′-CUU ACG CUG AGU ACU UCG A-3′), siIGFBP5 (5′-GCA GAU CUG UGA AUA UGA A-3′) and siSTAT3 (5′-GAA UCA CGC CUU CUA CAG A-3′).