Figure 2: Loss of Activin-β/dSmad2 signalling results in hemocyte defects.
From: Regulation of Drosophila hematopoietic sites by Activin-β from active sensory neurons
(a–d) Loss of Actβ in neurons affects hemocytes, lateral view of larvae (top panels) with close-up (lower panels). (a) Control HmlΔ-DsRed /+ (b) Actβ mutant, HmlΔ-DsRed/+; ActβED80/ED80. (c) Actβ RNAi silencing with ubiquitous driver Actin5C-GAL4. (d) Actβ RNAi (UAS-Act RNAi Vienna GD) silencing in PNS neurons, md neuron specific driver 21-7-GAL4. (e) Hemocyte counts in segmental HPs (bracketed areas in a–d top panels), n=4–7 per genotype. (f) Fraction of circulating hemocytes, n=4 to 8 per genotype. (g) Total hemocyte counts per larva, n=4 to 8 per genotype. In e–g experiment numbers relative to matching control cohorts, genotypes HmlΔ-DsRed/+; ActβED80/ED80 (matching control: HmlΔ-DsRed/+); Actin5C-GAL4, HmlΔ-DsRed/+; UAS-Actβ RNAi/+ (matching control: Actin5C-GAL4, HmlΔ-DsRed/+); 21-7-GAL4, UAS-mCD8GFP, HmlΔ-DsRed/+; UAS-Actβ RNAi/+ (matching control: 21-7-GAL4, UAS-mCD8GFP, HmlΔ-DsRed/+). Hemocyte numbers were assessed in larvae 64–75 h AEL (2.2–2.6 mm) (h–j) Hemocyte-specific RNAi silencing of Actβ pathway components, driver HmlΔ-GAL4, UAS-GFP; He-GAL4. Top panels whole larva lateral view, lower panels closeups. (h) Control; (i) UAS-put RNAi; (j) UAS-dSmad2 RNAi. (k) Hemocyte counts in segmental HPs (bracketed areas h-j in top panels), n=4 to 7 per genotype. (l) Fraction of circulating hemocytes, n=7 to 10 per genotype. (m) Total hemocyte counts per larva, n=7 to 11 per genotype; larvae 53–60 h AEL (1.8–2.0 mm). In (k–m) experiment numbers relative to matching control cohorts are shown; genotypes HmlΔ-GAL4, UAS-GFP/+; He-GAL4/+ (control); HmlΔ-GAL4, UAS-GFP/+; He-GAL4/UAS-put RNAi or HmlΔ-GAL4, UAS-GFP/UAS-dSmad2 RNAi; He-GAL4/+ (experiment). Scale bars:, a–d and i,j 0.5 mm. Error bars represent s.d., and two-tailed t-test values correspond to NS (not significant) P>0.05; *P≤0.05; **P≤0.01; ***P≤0.001; ****P≤0.0001.
