Transfection and siRNAs

Oligofectamine was used for siRNA silencing in HeLa, H1299, or 293 T cells, and DharmaFECT1 was used for the siRNA silencing in mouse ESCs and F9 cells, whereas Lipofectamine 2000 was used for transient transfection as described previously^25,26,41. Typically, 50 nM of each siRNA or their combinations were transfected into target cells for 48 h and cells were directly lysed in SDS or NP40 lysis buffers⁴¹. For verification of the effects, usually, two or three independent siRNAs were designed to examine the knockdown efficiency and the consequences of knockdown on target proteins⁴¹. The siRNAs for human genes are: LSD1-1: GGAAGAAGAUAGUGAAAAC; LSD1-2: AGUGAAAACUCAGGAAGAA; LSD1-3’UTR: GGGAGGAACUUGUCCAUUA; DCAF5-1: GCUGCAGAAACCUCUACAA; DCAF5-2: AUCACCAACUUCUGACAUA; L3MBTL3-1: GAUGCAGAUUCUCCUGAUA; L3MBTL3-2: GGUACCAACUGCUCAAGAA; SMARCC1: GGAACAAAGUGUCGGAACA; SMARCC1-2: GGAUGAAUGAGGAGGAUUA; SMARCC2-1: GCACAGACAUGUACACAAA; SMARCC2-2: GGAUGAGGAGAAAGG-GAAA; SMARCB1-1: ACGCUGAGAUGGAGAAGAA; SMARCB1-2: ACACUAAGGAUC-ACGGAUA; SET7-1: GGGCAGUAUAAAGAUAACA; SET7-2: ON-TARGETplus Human SETD7 (80854) siRNA-SMARTpool. The siRNAs for mouse genes are: mLSD1-2: AAGGA-AAGCTAGAAGAAAA; mL3MBTL3-1: GCTGAGGTTTGTGGATATA; mL3MBTL3-2: GCUCGAGGCUGCAGACAAA; mSMARCC1-1: GGAUGAAUGAAGAGGAUUA; mSMARCC1-2: GCUAACAAGUUGAAGAUA; mouse LSD1-1 is the same as human LSD1-1, and control siRNA for luciferase: CATTCTATCCTCTAGAGGA. All siRNAs were synthesized from Horizon Discovery.