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. 2014 Sep 4;289(43):29937–29947. doi: 10.1074/jbc.M114.573659

FIGURE 3.

FIGURE 3.

LSD1 is required for SREBP1-mediated transcription in mammalian cells. A–C and E, luciferase reporter assays detect the effects of LSD1 knockdown in HEK293 cells using lentivirus-based shRNA on SREBP-1a-induced activation of a basal promoter that contains three tandem SRE sites (A) or the 1-kb human FAS gene promoter (B), SREBP-1c-induced activation of the 1-kb human FAS gene promoter (C), and SREBP-1a or SREBP-2 -induced activation of the human LDLR gene promoter (E). D, effects of LSD1 knockdown and insulin (200 nm, 18 h) on the FAS promoter activity in HEK293 cells. F, ChIP-qPCR shows the effects of LSD1 knockdown on HA-tagged nuclear SREBP-1a binding to the FAS promoter in HEK293 cells. HA-tagged G4DBD-VP16-TAD and 2xGBE-Luc were co-transfected as the internal control. G, immunostaining for overexpressed Flag-nSREBP-1a (amino acids 1–490) when co-transfected with LSD1-shRNA or NS-shRNA (control) in HEK293 cells. The nuclei were visualized with DAPI staining. The data represent means ± S.D. (n = 3). *, p < 0.05; #, p < 0.01 versus NS; §, p < 0.05; clover symbol, p < 0.01 versus vector.