Regenerative Inflammation in Drosophila

To measure the regenerative inflammation index, we need to measure the number of mitotic cells per midgut and the relative expression of regenerative inflammation genes with or wiout dietary, probiotic or pharmaceutical interventions. Midgut stem cell mitosis will be measured via immunohistochemistry using an antibody against phosphorylated histone 3 (anti-pH3; Cell Signaling), as previously described [Apidianakis, 2009]. For relative gene expression we will perform RT-qPCR using total RNA from 20 midguts (in biological triplicates) to assess the expression of Dl/DLL1, upd3/IL-6, vein/EGF and eiger/TNF genes, normalized to the expression levels of the reference genes tub, gapdh and rpl32 using the 2-ΔΔCt method.