Figure 3: Pellino regulates Toll-mediated immune responses in vivo.

(a) The UAS-GAL4 strategy was used to test the in vivo function of pellino. Ubiquitous knockdown of pellino by the Tub-gal4 resulted in animal lethality at the late larval stage. (b,c) Flies containing overexpression or knockdown of pellino specifically in fat bodies by the Cg-gal4 driver were lysed to measure expression levels of pellino mRNA (b) or protein (c). Error bars represent s.d. (n=3). (d,e) Pellino knockdown, MyD88 knockdown (positive control) and Cg-gal4 flies (wild-type control) were infected with M. luteus for 12 h and lysed for quantitative RT-PCR analysis to measure mRNA levels of drosomycin (d) and metchnikowin (e). Error bars represent s.d. (n=3). (f,g) Pellino overexpression flies and controls were infected with M. luteus for 12 h and then lysed for quantitative RT-PCR analysis to measure mRNA levels of drosomycin (f) and metchnikowin (g). Error bars represent s.d. (n=3). (h,i) Pellino knockdown (h) or overexpression (i) flies and controls were pricked with a needle previously dipped into M. luteus. Bacterial replication was monitored at different times as indicated. (j,k) Pellino knockdown (j) or overexpression (k) flies and controls were infected with M. luteus and monitored daily for mortality. Error bars represent s.d. (n=3). For data from b and d–g, the two-tailed Student’s t-test was used to analyse statistical significance. For data from h and i, the rANOVA test was used to analyse the variance of repeat measures. For data from j and k, the Log-rank test was used to analyse the variance of the survival rates between two groups. *P<0.05, ***P<0.001, versus control groups.