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Fig. 6 RNA-sequencing identifies novel DEGs that distinguish CTRL from EAE-DV ventricles during larval and adult stages. A. Schematic outlining the RNA-sequencing and qPCR validation pipeline. B. A Venn diagram depicting unique and overlapping upregulated and downregulated DEGs relative to CTRLs between EAE-DV males and females. C. qRT?PCR on pooled isolated 5 mpf male and female ventricles reveals reproducibly dysregulated genes (n = pools of 3 males, >6 females per condition). D. qRT?PCR on pooled isolated 1-year-old male ventricles show that slc25a33, ankrd9, dusp2, and eya4 are significantly decreased in individuals with a moderate reduction in E/A ratio relative to those with a severe decrease in E/A ratio (n > 8). Fold change is calculated relative to the average normalized gene expression level of all sampled individuals. E. qRT?PCR on pooled isolated 20 dpf ventricles identifies a subset of the adult DEGs that are dysregulated during larval stages (n = 6). For C?E, statistical significance was determined using an unpaired, two-tailed Student's t-test assuming equal variances for groups with a normal distribution or a nonparametric Mann?Whitney test for comparisons between groups without normal distribution. For E, outliers were excluded using ROUT (Q = 1%). P-values are shown. ns, non-significant (P > 0.05).