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. 2021 May 6;16(5):e0251184. doi: 10.1371/journal.pone.0251184

Fig 11. Transgenic expression of a VPS24 mutant lacking the C-terminus disrupts VPS24 function and reveals colocalization of VPS24 with VPS28 at the expanded, ubiquitin-positive lysosomal compartment.

Fig 11

(A) Schematic representations of control (WT VPS24-FLAG) and C-terminal truncation mutant (VPS24-FLAG 1–178) VSP24 proteins carrying a C-terminal FLAG tag. The MIM1 [Microtubule-interacting and transport (MIT)-interacting motif 1] domain present in the VPS24 C-terminal mediates binding interactions with MIT domain proteins such as VPS4. The vps24 mutant form lacks this interaction domain. (B) Western blot analysis of transgene products following neuronal expression of the control or mutant transgene. Lysate was prepared from fly heads and Tubulin (TUB) was used as a loading control. (C-F) Confocal immunofluorescence images of CNS neurons following neuronal transgene expression in a WT or vps24 mutant background. (C) Neuronal expression of WT VPS24, but not VPS24 1–178, rescues the neuronal phenotype of the vps24 mutant. (D) Neuronal expression of VPS24 1–178, but not WT VPS24, in a WT background mimics the neuronal phenotype of the vps24 mutant. (E) Colocalization of VPS24 1–178 and endogenous VPS28 following neuronal transgene expression in a WT background. (F) Colocalization of VPS24 1–178 and the expanded lysosome membrane compartment following neuronal transgene expression in a WT background.