Extended Data Fig. 3: hnRNPA2B1 and MBNL1 mediate the alternative splicing of CARNS2 under hypoxia.
From: Carnosine regulation of intracellular pH homeostasis promotes lysosome-dependent tumor immunoevasion
a, Immunoblotting analysis of MBNL1 expression in whole-cell lysates (WCL) and nucleus lysates from PLC cells with normoxia or hypoxia treatment for 48 h. b, RNA immunoprecipitation analysis of the binding of endogenous CARNS mRNAs by MBNL in PLC cells. Cells were treated under normoxia or hypoxia for 24 h (n = 3 biological replicates). c, Venn diagram showing overlapping proteins bound to both MBNL1 protein and CARNS pre-mRNA in PLC cells. d, qPCR and Immunoblotting analysis of hnRNPA2B1 mRNA levels (n = 3 biological replicates) (left panel) and protein levels (right panel) in PLC cells expressing NTC, shHIF-1α or shHIF-2α, cultured under normoxia or hypoxia for 24 h. e, Binding of the endogenous CARNS mRNAs by HnRNPA2B1 in PLC cells expressing NTC and shMBNL1 was determined by RNA immunoprecipitation. Cells were treated under normoxia or hypoxia for 24 h (n = 3 biological replicates). Data presented as mean ± s.d. (b, d, e). Statistical significance was determined by two-tailed unpaired Student’s t-test (b, d, e).
