FBXO11 activates erythroid gene expression by degrading BAHD1. (A) WT and FBXO11 KO (clone F11KO #2; supplemental Figure 2C) HUDEP-2 cells were treated with Cas9 plus BAHD1 or control (Ctrl) sgRNAs and analyzed for erythroid markers after 5 days of induced maturation. The graph shows the mean ± SEM from 3 biological replicate experiments. (B) Cell pellets from FBXO11 KO HUDEP-2 cells that were treated with Cas9 + BAHD1 or Ctrl sgRNAs after 5 days of induced maturation. (C) WT and F11KO HUDEP-2 cells expressing BAHD1 or Ctrl shRNAs were analyzed as described for panel A. (D) Comparison of transcriptomes in F11KO HUDEP-2 cells expressing BAHD1 vs control (Ctrl) shRNA. The graph at left shows differential gene expression as described for Figure 3A. The right panels show the results of GSEA. (E) Heat map showing erythroid genes whose expression was decreased in F11KO vs WT HUDEP-2 cells (>twofold; false discovery rate [FDR], <0.05) and upregulated by the expression of BAHD1 shRNA (B1-sh1) in F11KO cells (>twofold; FDR, <0.1). *P < .05; **P < .01; ***P < .001 (unpaired Student t test).