Fig. 4

Stimulated microglia morphology changes upon treatment with MSC-EVs. (A) Representative images of C20 microglia cells in -CTL group (unstimulated), +CTL group (stimulated with 5ng/mL each of IFN-γ and TNF-α) and + CTL + EVs (stimulated with 5ng/mL each of IFN-γ and TNF-α + treated with MSC-EVs). Actin, Golgi & plasma membrane (PM) (Red, Phalloidin/WGA), Nuclei (Blue, Hoechst), Scale bars = 100 μm (columns 1–3 images) and 50 μm (column 4 images). (B) Individual min-max normalized cellular features (i) Perimeter, (ii) Major Axis Length and (iii) Form Factor. (iv)Normalized principal component 1(PC1_morpho) calculated using 21 min-max normalized features represents 72.2% of the variance. Each point is a median of ~ 700–1000 cells/well. Bar indicates the mean of the 8 wells/group. *p < 0.001 vs. + CTL for all groups conducted using Brown-Forsythe and Welch one-way ANOVA with multiple corrections using Dunnett T3 testing