LSD1 cooperates with GATA1 to promote glycolysis. (A) ChIP-sequencing (ChIP-seq) analysis. Distribution of LSD1-enriched peaks in HEL cells, relative to annotated regions in the genome. (B) Motif analysis of 8280 LSD1-enriched peaks. Motif and statistical analyses were performed by using HOMER software. (C) Coimmunoprecipitation of LSD1 and GATA1. Input lane contains the 10% of the total amount of whole cell extract, relative to IP lanes. (D) Gene expression changes in GATA1-KD HEL cells. (E) Decreased GLUT1 protein in GATA1-KD HEL cells. Samples were collected at indicated days after doxycycline (DOX) induction. (F) Reduction of glucose uptake in GATA1-KD HEL cells, as determined by 2-NBDG incorporation. (G) Reduction of glycolytic activity in GATA1-KD HEL cells. Values are mean ± standard deviation of 5 assays. (H) LSD1-dependent enrichment of GATA1 at ALAS2 and GLUT1 gene enhancers. ChIP-qPCR analyses were performed in control (ctrl) and LSD1-KD HEL cells. Enrichment values were calculated as percentage of input DNA. (I) Growth curve of ctrl and GATA1-KD HEL cells. (J) Downregulation of GATA1 protein in LSD1-KD HEL cells. Samples were collected at indicated days after DOX induction. (K) Effect of proteasome inhibition on GATA1 protein in LSD1-KD cells. The cells were treated with MG132 (10 µM) for 6 hours before harvest. Values are mean ± standard deviation of triplicate results except for those in panel G. *P < .05, **P < .01. 5′UTR, 5′ untranslated region; IgG, immunoglobulin G; TSS, transcriptional start site; TTS: transcriptional termination site.