Extended Data Fig. 9: B-isox affinity chromatography evidence of ACTC1 associated with R3Gs in HMZ myocardium and R3G antagonized SG assembly in U2OS cells.
From: Dysregulated ribonucleoprotein granules promote cardiomyopathy in RBM20 gene-edited pigs
a, B-isox affinity chromatography proteomics demonstrating cardiomyopathy gene enrichment analysis and specifically ACTC1 pulldown from R3G-laden HMZ gene-edited pig myocardium, see (Fig. 4d, e). Repeated three times, this experiment consistently produced similar results. Proteomics statistical analysis as described in (Fig. 4d legend). b, Double IHC for RBM20 and ACTC1 in 7-day HMZ pig myocardium in transverse section confirming reciprocal staining intensity of R3Gs and ACTC1, see (Fig. 6a) for analogous longitudinal sections. This myocardial IHC experiment repeated in three different HMZ piglets produced identical results. Scale bar, μm. c, GFP signal and SG marker, G3BP1, by ICC with DAPI in NaAsO2 treated GFP-RBM20-R636S transfected U2OS cells, demonstrating that cells expressing R3Gs fail to construct G3BP1 SGs, consistent R3G sequestration of G3BP1, a protein critical for SG assembly. Similar results observed in multiple independent U2OS cell transfection ICC experiments. Scale bars, μm.
