Figure 3: LSD1 controls OXPHOS through Nrf1.
From: LSD1 promotes oxidative metabolism of white adipose tissue
(a) HOMER motif analysis of LSD1 ChIP-seq data unravel Nrf1 binding sites among the top scoring motifs. (b) Cell lysates from 3T3-L1(Ctrl) and 3T3-L1(LSD1) differentiated adipocytes were subjected to sucrose gradient centrifugation. Aliquots of the gradient fractions and total cell lysate (Input) were subjected to immunoblot analysis for LSD1 and Nrf1. (c) Co-immunoprecipitation of LSD1 and Nrf1 in differentiated 3T3-L1(LSD1) cells. Western blots were probed with the indicated antibodies. (d) Western blot analysis of LSD1 and Nrf1 expression in 3T3-L1(Ctrl) and 3T3-L1(LSD1) cells at indicated days. β-Actin served as a loading control. (e) Venn diagram depicting the number of genes with LSD1 and Nrf1 peaks at the promoter in differentiated 3T3-L1(Ctrl) cells (day 7). (f) Enriched pathways obtained from GO term analyses for the indicated sets of genes. (g) Relative transcript levels of the indicated markers in differentiated C3H-10T1/2(LSD1) cells transfected with unrelated control siRNA (siRNA Ctrl) or siRNA directed against mouse Nrf1 5′ untranslated region (siRNA mNrf1) and infected with inducible empty control vector (Ctrlind) or human NRF1 (hNRF1ind) lentivirus, treated with Doxycycline (Dox) at day 3 of differentiation. (b–d) and (g): n=9; s.d. represents+s.e.m. Experiment (g) was independently repeated at least three times in triplicate. Statistical analysis was performed using (a) HOMER analysis or (g) two-tailed Student’s t-test. *P<0.05 and ***P<0.001.
