Figure 4

Western blot and RNA-seq analyses confirm downregulation of ARHGAP19 as well as cell cycle, motor, and muscular cytoskeleton pathways.

(A and B) Protein expression levels of ARHGAP19 in iPSc-derived SMN lines (A) and patient-derived fibroblasts (B). Fibroblasts from P5 (F5-II:2), P6 (F6-II:7), and P11 (F10-II:1) harboring the c.419G>A (p.Gly140Asp), c.203T>C (p.Leu68Pro), and c.85A>G (p.Asn29Asp) variants, respectively, were analyzed by Western blot, as well as fibroblast-derived iPSc SMNs from P5 (F5-II:2) and P6 (F6-II:7) patients. (C) Volcano plot showing log₂ of fold change in ARHGAP19 mutants compared with controls and –log10 (adjusted P-value). (D) Pathway enrichment: differentially expressed genes with P value < 0.005 were selected for performing pathways enrichment analysis. Top 10 pathways with the highest enrichment scores were plotted, showing 3 cellular pathways highly enriched with dozens of affected genes and a very low P-value: cell cycle, motor proteins, and cytoskeleton in muscle cells. (E) Pathway scores per sample: enriched pathways were analyzed to compare their expression levels in patients and controls. Some pathways are overexpressed in patients while most of them are downregulated.