Extended Data Fig. 8: Role of Osbpl9 and Pla2g6 isoforms in lipid metabolism.
From: Liver RBFOX2 regulates cholesterol homeostasis via Scarb1 alternative splicing in mice
A.- Capillary electrophoresis and quantification of percentage splice in (PSI) for Osbpl9 (exon 6), in LWT and LΔRbfox2 hepatocytes upon treatment with SSO11.1 or Scr control. Statistical significance was determined by one-way ANOVA with Tukey’s multiple comparisons test of biologically independent samples (n = 5-6). B.- Heatmap showing LC-MS metabolomic analysis of LWT and LΔRbfox2 hepatocytes treated with SSO11.1 or Scr. C.- Western blot showing PLA2G6 expression in wild type and RBFOX2-deficient hepatocytes (top). Cryo-EM maps of PLA2G6 dimers showing tight interaction of the catalytic domains (orange) of each monomer with ankyrin repeats (purple) oriented outward from the core. Ankyrin repeats face ‘claw-like’ towards membrane phospholipids. Insert: 90-degree rotation of the structure detailing the region corresponding to exon 10 of Pla2g6L, a 55-amino intrinsically disordered proline-rich region at the interface of the ankyrin repeats and the catalytic domain. D.- Diagram showing design of splice-switching oligos targeting Pla2g6 alternative splicing at exon 10. SSO5.1 is designed to promote exon skipping (top) as validated by semi-quantitative PCR analysis (bottom). E.- Quantification of the effect of SSO5.1 on the described lipid species as determined by LC-MS normalised to internal standard and total cell counts. Results are represented as mean ± s.e.m. (n = 5-6). Statistical significance was determined by two-sided unpaired t-test of biologically independent samples.
