Fig. 7: PTBP1 regulates the alternative splicing of Arrb1 to control endothelial cell migration.
From: Endothelial deletion of PTBP1 disrupts ventricular chamber development
a Representative images of the Sashimi plots showing the exon splicing of Arrb1. The Sashimi plots show the densities of exon-including and exon-skipping reads as determined by rMATS-turbo analysis. b Gel electrophoresis analysis of the ARRB1 isoforms in endothelial cells from Nfatc1-Cre;Ptbp1fl/fl, Tie2-Cre;Ptbp1fl/fl and Cdh5-CreERT2;Ptbp1fl/fl hearts. The values indicate expression ratios of the short ARRB1 isoform (ARRB1-S, lacking exon 13) and the long ARRB1 isoform (ARRB1-L, including exon 13) (S/L). c qRT-PCR analysis of ARRB1 from RNA immunoprecipitation assay of HUVECs using anti-PTBP1. RNA enrichment is determined relative to the nontargeting IgG control. RIP assay was described in the schematic diagram. n = 3 independent experiments. d AONs transfection reverses the switching change of S/L ratio caused by PTBP1 knockdown (LV-sh-PTBP1). The S/L values indicate ratios of ARRB1-S to ARRB1-L. AONs-1 and AONs-2 represent different oligonucleotide sequences e, f AONs rescued the decreased HUVEC migration caused by PTBP1 knockdown (LV-sh-PTBP1). n = 3 independent experiments. Scale bars, 200 µm. LV-sh-PTBP1, lentivirus-shRNA-PTBP1; AONs, antisense oligonucleotides; Ctrl, control lentivirus-shRNA or control AONs. g, h ARRB1-L overexpression (oe-ARRB1-L) or ARRB1-S overexpression (oe-ARRB1-S) could promote the HUVEC migration. n = 3 independent experiments. Scale bars, 200 µm. Ctrl control lentivirus-vector. The data are presented as the mean ± s.e.m. P values were calculated by unpaired two-tailed t test. Source data are provided as a Source Data file.
