Figure 3.

Genes required for efficient bacterial engulfment in adult Drosophila. (A) Quantitative RT–PCR confirms reduced expression of nimC1 and eater transcripts in young female flies of the genotypes He-Gal4, UAS-GFP; UAS-RNAi nimC1 (left) or eater (middle). Two biological replicates are shown. He-Gal4; UAS-GFP, UAS-eater flies showed increased gene expression and served as a positive control (right). Fold changes were calculated using △△Ct values relative to the reference gene RpL32 (rp49) and the control genotype He-Gal4, UAS-GFP, using multiple technical replicates for each biological replicate. (B–C) Box plots of phagocytic ability, measured as the number of fluorescent Escherichia coli per active hemocyte at 90 min postinjection. Boxes represent the middle two quartiles, separated by a line representing the median, and the whiskers show the 10th and 90th percentiles. Dots indicate outliers. A dashed line marks the mean value of events per cell. N = number of flies and (total cells) assayed. (B) Reduced function of eater or nimC1 reduces bacterial uptake by adult hemocytes. The following genotypes were compared: He-Gal4, UAS-GFP, UAS-RNAi eater and He-Gal4, UAS-GFP, UAS-RNAi nimC1. The isogenic control strain (He-Gal4) showed significantly greater phagocytic ability compared with either knockdown line (P < 0.05). (C) Increased Rab5 increases the number of phagocytic events per hemocyte. There is a significant difference between He-Gal4 outcrossed to UAS-GFP control vs. crossed to UAS-Rab5 wt (P < 0.0001). The parental control strain (He-Gal4, UAS-GFP) is also shown.