Fig. 3: Oxygen-dependent reconfiguration of RBP-regulated cellular networks.
From: A network of RNA-binding proteins controls translation efficiency to activate anaerobic metabolism
a Hypoxia-induced rewiring of RBP-regulated processes by Gene Ontology (GO) pathway enrichment analysis. b GO analysis of representative enriched biological processes for downregulated proteins when indicated RBPs are silenced. c Protein output regulation of each glycolytic effector by the hypoxia-adaptive RBPs PCBP1, hnRNP A2/B1, and HuR, as determined by TMT-pSILAC (three independent experiments pooled into a single sample for measurement). Orange dotted line represents the MS regulatory threshold, validated empirically at the steady-state protein level by immunoblot, and at the translation efficiency level by qRT-PCR of ribosome density fractions. d Left panel: protein expression of glycolytic effectors are induced under hypoxic conditions (confirmed by TMT-pSILAC). Synergistic regulation of glycolytic proteins (right panel) by hypoxia-adaptive RBPs (middle panel) as determined by TMT-pSILAC analysis. Measurements of e glucose uptake and f lactate production in U87MG treated with indicated siRNAs (for 48 h prior to following experimentation). Color scheme: red, normoxia; blue, hypoxia. NS: non-silencing. Asterisk denotes statistical significance calculated using two-sided Student’s t-tests compared to NS control. Exact p values e: PCBP1 siRNA (p = 0.04), hnRNP A2/B1 siRNA (p = 0.04), PCBP1 + hnRNPA2/B1 siRNA (p = 0.02), eIF5B siRNA (p = 0.02). Exact p values f: PCBP1 siRNA (p = 0.01), hnRNP A2/B1 siRNA (p = 0.03), PCBP1 + hnRNPA2/B1 siRNA (p = 0.02), HuR siRNA: (p = 0.03), eIF5B siRNA (p = 0.04). Data represent mean ± SEM (error bars) of three independent experiments. Source data are provided as a Source data file.
