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. 2022 Dec 4;11(23):3923. doi: 10.3390/cells11233923

Figure 4.

Figure 4

MSC-derived EVs do not affect the NLRP3-inflammasome, nor the p65 pathway in patients-derived iAstrocytes. (a). Representative WB immunoreactive bands for NLRP3 and P-p65 in patient-derived iAstrocytes differentiated from control (CTR) subjects, C9orf72, and SOD1 patients treated or not 24 h with human MSC-derived EVs. (b,c) Quantitative representation of WB densitometric expression signals for (b) NLRP3 and (c) P-p65 normalized to GAPDH. EV treatments did not show any significant effect on protein expression. Data are presented as means ± SEM of n = 3 independent experiments per patient. One control and 2 patients per group are included (One-way ANOVA, followed by Bonferroni post hoc test). (d–f) ELISA quantification of cytokines, reported as pg/mL, for (d) IL-1β, (e) IL-6 and (f) CCL2, in the conditioned media of iAstrocytes differentiated from control CTR, C9orf72, and SOD1 subjects treated or not 24 h with human MSC-derived EVs. Data are presented as means ± SEM of independent experiments as follows: n = 3–6 for one CTR, n = 4–6 for 2 C9 donors and n = 5–6 for 2 SOD1 donors; statistical significance for p < 0.05 at least (* p < 0.05 vs. untreated iAstrocytes; F(2,30) = 8.936, F(2,24) = 2.833, F(2,30) = 2.677, for IL-1β, IL6 and CCL2, respectively, in untreated vs. EV-treated iAstrocytes; two-way ANOVA with Sidak’s multi comparison test).