Fig. 1: Inflammatory features of murine dysferlinopathic skeletal muscles.
A Volcano plot analysis of genes differentially expressed in 6 m BlAJ vs 12 m BlAJ muscles: red and yellow dots represent genes upregulated in 6 m BlAJ and 12 m BlAJ, respectively, with a p value < 0.05 (corresponding to a -Log10(p value) > 1.30). B Gene ontology (GO) pathways enrichment in 6 m BlAJ and 12 m BlAJ mice, or both. For GO analysis 837 genes upregulated in 6 m BlAJ, 755 gene upregulated in 12 m BlAJ and 10,638 non-differentially expressed genes (based on RNA-seq data) were used. Only significant pathways (p value < 0.05) are shown. Redundant pathways are omitted from the figure. C Volcano plot analysis of genes involved in complement activation (Biological process; GO: 0006956) and complement receptor activity (Molecular function; GO: 000487). Red and yellow dots represent genes upregulated in 6 m BlAJ and 12 m BlAJ, respectively, with a p value < 0.05 (corresponding to a -Log10(p value) > 1.30). D GSEA plot for the annotated dataset “Hallmark_Complement” by the Molecular Signatures Database (MSigDB). Geneset size: 143; Normalized enrichment score (NES): 1.43; False discovery rate (FDR) q value: 0.064. E WB expression of IP subunits and inflammatory mediators in psoas muscle of 12 mo BlAJ and 12 mo C57Bl mice. F Evaluation of CD45+ cells, CD4+ and CD8+ T-lymphocytes and Tregs in skeletal muscles of 12 m BlAJ and C57Bl mice. G Representative FACS of CD4 and CD8 cell subpopulation profiles. The numbers within the panels indicate the percentage of each population of live cells, a gate of CD45-positive events defined infiltrating inflammatory cells. Within the CD45+ cells, two populations were separated by the expression levels of CD4 or CD8, whose subpopulations were separated according to CD44 and CD62L.Within CD4+ cell population, positivity for CD25 was also determined. H Evaluation of T-cell subsets—naive, central memory (CM), effector memory (EM)—expressing CD4 and/or CD8 in the muscles of 12 m C57Bl and 12 m BlAJ. Data are presented as mean ± SD of n = 3 independent experiments with n = 3–12 animals/group. One-way ANOVA and Student t-test: *p < 0.05, **p < 0.01, ***p < 0.001 and ****p < 0.0001.
