Extended Data Fig. 10: 3’UTR splicing-mediated cytoplasmic localization enhances CTNNB1 expression.
From: Pan-cancer pervasive upregulation of 3′ UTR splicing drives tumourigenesis
a,b, Effect of overexpressing CTNNB1 CDS, CDS + 3’FL and CDS + 3’SP on endogenous CTNNB1 transcript (n = 3 independent experiments) (a) and exogenous CTNNB1 protein (b) expression in DLD-1. c, Luciferase activity of reporter constructs with CTNNB1 3’FL and 3’SP in DLD-1 (n = 3 independent experiments). d, Effect of actinomycin D (ActD) treatment on the transcript levels of exogenously expressed CTNNB1 CDS, CDS + 3’FL and CDS + 3’SP in Hep3B and SNU398 (n = 3 independent experiments). e,f, Effect of cycloheximide (CHX) (e) or MG132 (f) treatment on exogenously expressed CTNNB1 protein levels of in Hep3B and SNU398. g, Subcellular distribution of CHEK1 3’FL and 3’SP transcripts following nuclear-cytoplasmic fractionation of Hep3B and SNU398 cells (n = 3 independent experiments). MALAT1 was used as a nuclear control. The 3’FL:3’SP transcript ratios in each cellular compartment are shown in the table below. h, RNA-FISH showing transcript localization of CHEK1 3’FL and 3’SP in SNU398. EV: empty vector; CDS: coding sequence; 3’FL: full length 3’UTR; 3’SP spliced 3’UTR. a,c,d,g, Mean ± SEM; unpaired Student’s t-test *p < 0.05, **p < 0.01, ***p < 0.001. b,e,f,h, Data shown represent three independent experiments.
