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Fig. 6 | Parasites & Vectors

Fig. 6

From: Eimeria falciformis secretes extracellular vesicles to modulate proinflammatory response during interaction with mouse intestinal epithelial cells

Fig. 6

Eimeria falciformis sporozoite EVs upregulate caspase 11 and NLRP6 inflammasomes and release LDH. a, b MIECs (1 × 106) were plated and stimulated with 50 µg of EfSEVs for 6, 12, 18 h and 10, 30, 50 and 100 µg of EfSEVs for 24 h. An equivalent volume of PBS (0 µg of EfSEVs) was used as experimental control for the dose treatment and 0 h was used as control for the time-dependent treatment. An equal volume of NC particles was set as a negative control. RNAs were extracted and analyzed by qPCR for caspase 11 and NLRP6 mRNA expression. Bars represent mean ± standard deviation values in triplicate experiments. The difference in mRNA expression was compared with control treatments and considered to be significant at *P < 0.05, **P < 0.001, ***P < 0.001. c EfSEVs induced secretion of LDH in MIECs. 1 × 106 MIECs were stimulated with increasing doses (10, 20, 30, 50, 100 µg) of EfSEVs. Also, 50 µg of EfSEVs was used to stimulate MIECs at increasing duration (0, 6, 12, 18, 24 h). PBS (0 µg of EfSEVs) was used as experimental control for the dose treatment and 0 h as the control for time-dependent treatment. An equal volume of NC particles was set as a negative control in both cases. MIECs were harvested at the end of the treatment and assayed for LDH production using the Chekine™ LDH assay kit in triplicate. Differences in the time- and dose-dependent release of LDH were compared with the control treatments and considered to be significant at *P < 0.05, **P < 0.001, ***P < 0.001. Abbreviations: LDH, Lactate dehydrogenase; NLRP6, nucleotide-oligomerization domain (NOD)-like receptor pyrin 6

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