Fig 5. ird1 loss-of-function prevents normal crystal cell development and activates Toll signaling in the fat body.

A. Crystal cells in the posterior ends of control (+) and ird1^-/- larvae are visualized (arrowheads) by heat treatment (10 min, 70°C) and quantified. The plots indicate medians, quartiles and outliers for the number of black spots counted in images taken of the dorsal side of eight individuals per genotype. The significance level was estimated by Mann-Whitney U exact test (two-tailed). B. Hemocytes from control (+) and ird1^-/- larvae stained with the C1 antibody (green), which is specific for the crystalline inclusions in crystal cells (arrowheads), and Hoechst nuclear dye (blue). The plasmatocyte-specific eater-DsRed reporter (red) is never seen in cells that stain with the crystal cell-specific antibody (arrows). C. The percentage of larvae with visible black capsules 27–29 hours after infection with the parasitoid wasp Leptopilina boulardi. The graph shows the average of two independent experiments with at least 100 larvae for each genotype and trial. D, E. Toll signaling activity in larval fat bodies (D) and extracted hemocytes (E), visualized with the Drs-GFP reporter in wild-type control (+), Tl^10b heterozygotes (Tl^10b/+), ird1 null mutant larvae (ird1^-/-) or animals expressing UAS-ird1^IR (ird1^IR) in both fat body and hemocytes with two copies of the Cg-GAL4 driver (2xCg>). Values below the images in D indicate the proportion of individuals showing fat body-specific Drs-GFP (not counting spontaneous Toll-independent activation of Drs-GFP in the trachea).