Fig. 1

Knockdown of the OXPHOS complex genes in hemocytes affects the mitochondrial membrane potential, causes melanotic nodules and enhances the immune competence. A A schematic representation of oxidative phosphorylation (OXPHOS) complexes and the complex-specific knockdown target genes: ND-75 (NADH dehydrogenase (ubiquinone) 75 kDa subunit); SdhD (Succinate dehydrogenase, subunit D); ox (oxen); UQCR-C1 (Ubiquinol-cytochrome c reductase core protein 1); COX5B (Cytochrome c oxidase subunit 5B); ATPsynCF6 (ATP synthase, coupling factor 6). B Mitochondrial membrane potential was measured as a ratio of the MitoProbe™ TMRM signal intensity detected in the OXPHOS knockdown plasmatocytes to the signal detected in control plasmatocytes (n = 3, 2000–3500 eater-GFP-positive plasmatocytes per replicate). The data were analyzed using one sample t-test. C Examples of melanotic nodules found from hemocyte-targeted cV knockdown animals. Nodules are marked with arrowheads. Scale bars 500 µm. D Quantification of melanotic nodules detected in the hemocyte-targeted OXPHOS gene knockdown larvae (n = 100). * = 1% of the driverless background control larvae had melanotic nodules. E Mean percentage of melanized wasp eggs/larvae found in the controls and in larvae with OXPHOS knockdown in hemocytes (n = 150). E’ Melanization response to wasp infection in the animals with ND-75 and ATPsynCF6 knockdown in the fat body (n = 150). The data were analyzed using logistic regression with a binomial distribution with replication as a random factor. ns = not significant, * p < 0.05, ** p < 0.01, *** p < 0.001. (Schematic Fig. 1A modified from [10])