mtm and class II Pi3K68D have opposing roles for endolysosome homeostasis, distinct from class III Vps34. LysoTracker, red; DAPI, blue; shown for single cell. (A) Rescue of endolysosomal size upon mtm, Pi3K68D coRNAi in Kc₁₆₇ cells. (B, bottom) In hemocytes, class II PI3K depletion (IRPi3K68D) rescued mtm-dependent endolysosome size. Class III PI3K disruption alone (Vps34-KD, kinase dead, or vps34^Δm22) resulted in diffuse LysoTracker staining (top) rescued by mtm disruption. (C–D’) Size of individual LysoTracker-positive organelles upon single RNAi (gray) and mtm coRNAi (black) in Kc₁₆₇ cells or hemocytes (D and D’; Vps34^Δ unquantifiable). (E) Pi3K68D:GFP expression (Pxn-GAL4) led to enlarged endolysosomes, whereas Vps34-WT had little effect. Error bars indicate SEM.