Fig 10. Knockdown of ESCRT proteins suppresses the accumulation of ubiquitinated protein aggregates and Ref(2)P in Gba1b mutants.

(A-F) RNAi constructs were expressed using the pan-neuronal driver elav-GAL4 in Gba1b mutants (Gba1b^ΔTT/Gba1b^ΔTT) and controls (Gba1b^rv/ Gba1b^rv). Homogenates were prepared from fly heads using 1% Triton X-100. Western blot analysis was performed on the Triton X-100–insoluble proteins using antibodies to ubiquitin (Ub) and Actin, and on the soluble fractions using antibodies to Ref(2)P and Actin. (A-B) Representative images and quantification of (A) ubiquitin and (B) Ref(2)P from flies with and without Mvb12 knockdown. (C-D) Representative images and quantification of (C) ubiquitin and (D) Ref(2)P from flies with and without knockdown of lsn. (E-F) Representative images and quantification of (E) ubiquitin and (F) Ref(2)P from flies with and without knockdown of CHMP2B. At least three independent experiments were performed. Error bars represent SEM. *p < 0.05 by Student t test.