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Fig. 4 Zebrafish embryo model. mpeg1-eGFP transgenic zebrafish larvae were injected with 7,500 EV/larva and 18 h after injection, eGFP+ macrophages were isolated using FACS. A Kaplan?Meier graph shows the percentage of survival up to 48 h post-injection with EVs isolated with the TFF system. B, C Changes in the expression of tnf and il6 in macrophages were quantified by qPCR, and data were normalized to the housekeeping gene eef1a. Statistical analysis was performed using one-way ANOVA followed by Bonferroni's post-hoc test. * shows a significant difference between Co and EVs. Data are presented as mean ± SD, and p < 0.05 was considered statistically significant (n = 3 biological replicates, 100?150 embryos/condition). Co: larvae injected with HEPES buffer, hpi: hours post-injection