Fig. 5: Y-33075 promotes alternative splicing of OGT and OGA.
From: Small molecule splicing modulators that disrupt O-GlcNAc homeostasis
a DNA melt curve analysis following RT-qPCR reactions using displayed primers from HCT116 cells treated as indicated for 24 h. One representative curve shown from three independent biological replicates. b– d Gel electrophoresis of RT-qPCR reactions using displayed primers from HCT116 cells treated as indicated for 24 h (n = 1). Secondary products generated by Y-33075 indicated with red asterisks. c– e Schematic of secondary products identified in (b) and (d). f Canonical (top) and alternative 5’ splicing (bottom) pathways for OGA. g Predicted splice acceptor and donor sites in OGA between exons 11 and 12. Borders of exon 11, DI2, exon 12, and sub-DI2 indicated. Sub-DI2 alternative 5’ splice site indicated with pink arrow. Figure generated using SpliceAI-Visual (https://mobidetails.chu-montpellier.fr/)47. h RT-qPCR analysis of transcript abundance in HCT116 cells treated as indicated for 24 h (n = 3). Data are represented as mean ± s.d. Two-tailed paired samples t-test with p-values computed based on ACTB-normalized Cq values shown. Data normalized to DMSO. i Gel electrophoresis of RT-qPCR reactions from HCT116 cells treated as indicated for 24 h (n = 1). Expected sub-DI2-containing products denoted by pink asterisks. ΔCq (vs. ACTB) values normalized to DMSO. For sequencing results, see Supplementary Fig. 11–14. Source data are provided as a Source Data file.
