Fig. 5.

Autophagy inhibition increases LPS induced IL-1β secretion from both Brown Swiss and Holstein Friesian macrophages.

3MA inhibits autophagy by blocking autophagosome formation via the inhibition of type III Phosphatidylinositol 3-kinases (PI-3 K), leading to a down-regulation of inflammaosome activation, and thus IL-1β production. To assess whether MØ of both breeds have the capability to respond with IL-1 production to inflammasome activation, MØ were derived from PBMC isolated from (A) HF and (B) BS and adhered overnight prior to inhibition of autophagy with 3MA (from 10 mM to 1.25 mM) in the presence of LPS (5 μg ml⁻¹) for 24 h. Supernatants were collected and analysed for bovine IL-1β by ELISA. Absorbance was measured using a SpectraMax M2 plate reader. All data were post analysed using GraphPad Prism using two-way non-repeated measures ANOVA with Bonferroni post-hoc testing.