Fig. 2

Knockdown of the OXPHOS genes in hemocytes induces immune cell activation. A Flow cytometry analysis of larval hemocytes using the plasmatocyte (eater-GFP, y-axis) and lamellocyte (msn-mCherry, x-axis) in vivo hemocyte reporters. While the control animals mainly had plasmatocytes (high GFP), the knockdown of various OXPHOS genes induced immune-activated hemocyte types, including activated plasmatocytes (high GFP, low mCherry) and lamellocytes (high mCherry intensity, marked in red), and occasionally also lamelloblasts (low GFP) and prelamellocytes (low GFP, low mCherry). B Heatmap (n = 30) showing the average total hemocyte and lamellocyte (lc) count in w^GD control and hemocyte-targeted OXPHOS knockdown samples. C–C’’’’’ Quantification of total hemocytes and hemocyte types classified based on eater-GFP and msn-mCherry expression when knocking down the OXPHOS genes (n = 30). Total = total circulating hemocyte count, pc = plasmatocytes, act pc = activated plasmatocytes, lb = lamelloblasts, pre lc = pre-lamellocytes, lc = lamellocytes. cIII gene = UQCR-C1. The data on hemocyte counts were analyzed using a generalized linear model with a negative binomial distribution. Error bars indicate standard error of the mean. Asterisks indicate the statistical difference between the OXPHOS knockdowns and the control. D Mitochondrial membrane potential in plasmatocytes of wasp-infected larvae and of larvae with hep^CA or Toll^10b overexpression in hemocytes, as a ratio to the MitoProbe™ TMRM signal intensity in the control. ns = not significant, * p < 0.05, ** p < 0.01, *** p < 0.001