Figure 4. Targeted RNAi depletion of Smn in Drosophila immune cells yields melanotic masses and reduced viability.
A) Fraction of larvae that display MMs. RNAi mediated knockdown of SMN was carried out using the Drosophila GAL4/UAS system to drive expression using UAS-SmnJF (P{TRiP.JF02057}attP2) together with the following GAL4 drivers: da, daughterless (da) for ubiquitous knockdown; C15 (a composite driver that includes elav- (embryonic lethal abnormal vision), sca- (scabrous) and BG57-GAL4 see (Budnik et al. 1996; Brusich et al. 2015) for knockdown in both neurons and muscles; and Cg (Collagen 4a1 gap), for knockdown in the fat body, hemocytes, and the larval lymph gland. OreR is the control strain. A plus sign (+) indicates a wild-type chromosome. B) Representative image of a wild-type control and MMs in a larva with SMN depleted in the fat body, hemocytes, and lymph gland (Cg-GAL4>UAS-SmnJF). C) Number of MMs per animal with and without SMN depletion, as in (A). SmnHM (P{TRiP.HMC03832}attP40) is an alternative UAS RNAi line that targets Smn. D and E) Fraction of larvae with MMs (D) and number of MMs per animal (E) using the hemocyte specific Gal4 driver Hml (Hemolectin) together with the UAS-SmnJF transgene. Control strains as per panel A.