WDR11 cooperates with Hh pathway to regulate gene expression

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WDR11 cooperates with Hh pathway to regulate gene expression WDR11 cooperates with Hh pathway to regulate gene expression Gene expression analyses of GN11 after 48‐h treatment with purmorphamine (Pur) or solvent dimethylformamide (DFM) by RT–qPCR. Pur significantly induced Ptch1, Gli3, GnRH1 and Wdr11, while Emx2 was unresponsive (P = 0.44; t = 0.87). In FNCB4‐hTERT, Pur significantly induced EMX1, PTCH1, GLI3, GNRH1 and WDR11 mRNA levels compared to DFM. ShRNA‐mediated WDR11‐targeted knockdown (57%) in FNCB4‐hTERT cells or Wdr11 KO in mouse led to significant reduction in the expression of these genes, compared to the control. Data are shown as means ± SEM from three independent experiments after unpaired Student's t‐test (*P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001).RT–qPCR analyses of FNCB4‐hTERT and MEFs with WDR11 KD/KO, respectively. Pur treatment for 48 h caused a significant upregulation of GLI3 and EMX1 in control FNCB4‐hTERT cells, which was dampened down in WDR11 KD cells (n = 3). Similarly, the expression of Gli3 and Ptch1 was significantly upregulated by Pur in WT (n = 7), but dampened down by Wdr11 KO MEFs (n = 5). Data are presented as means ± SEM from independent experiments after unpaired Student's t‐test (*P < 0.05; **P < 0.01; ****P < 0.0001).Overexpression of GLI1 and GLI3 in FNCB4‐hTERT significantly upregulated EMX1 mRNA compared to empty vector, while WDR11 remained unresponsive. Data represent means ± SEM of three independent experiments with one‐way ANOVA followed by Dunnett's post hoc test (****P < 0.0001).RT–qPCR analyses of GNRH1 in FNCB4‐hTERT cells transfected with WDR11, EMX1, GLI3FL or GLI3R expression constructs. EMX1 or GLI3FL alone showed no effects. Co‐transfection of EMX1/GLI3FL caused a moderate increase (37%) of GNRH1, while co‐transfection of WDR11/EMX1/GLI3FL significantly increased the expression (231%). GLI3R/EMX1 or WDR11/GLI3R/EMX1 did not show any significant response. Data represent means ± SEM of three independent experiments with one‐way ANOVA followed by Dunnett's post hoc test (**P < 0.01; ***P < 0.001). Source data are available online for this figure. Yeon‐Joo Kim et al. EMBO Rep. 2018;19:269-289 © as stated in the article, figure or figure legend