Figure 5.

DRD2 signaling activates hypoxia inducible factors in normoxic cultures. A, Correlation between DRD2 and 12042 genes from TCGA was determined by Pearson correlation coefficients. Forty-nine genes with coefficients >0.5 or <−0.5 and FDR <0.05 were selected. Genes fitting these parameters were then analyzed using Enrichr. Top hits are shown in the graph, with combined score and adjusted p value. B, PDX GBM cells were treated with DRD2 agonist (30 nm) for 1, 2, or 4 d and then probed for expression of HIF1α by immunoblot analysis. DMSO-treated cells as vehicle-treated control and cells exposed to hypoxic conditions (1.5% O₂) were used as a positive control for HIF expression, and β-actin was used as a loading control. C, Western blots were used to analyze the level of proteins in the VHL–PHD regulatory pathway after treatment with either DMSO or DRD2 agonist (20 or 30 nm). β-Actin was used as a loading control. D, PDX GBM6 were exposed to DRD2 agonist (30 nm) and treated with HIF inhibitor for 24, 48, and 72 h; SOX2, MMP2, and CMYC protein levels were assayed.