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. 2021 Feb 4;17(2):e1008859. doi: 10.1371/journal.pgen.1008859

Fig 4. Tissue-specific knockdown of nSMase in Gba1b mutants differentially alters ubiquitinated proteins and Ref(2)P.

Fig 4

(A-F) Neutral sphingomyelinase (nSMase)-RNAi was expressed using tissue-specific drivers in Gba1b mutants. Homogenates were prepared from fly heads and thoraces using 1% Triton X-100 lysis buffer. 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-D) Representative images and quantification of ubiquitin in (A) thoraces and (B) heads and Ref(2)P in (C) thoraces and (D) heads of flies with and without nSMase knockdown in flight muscle using the Act88F-GAL4 driver. (E,F) Representative images and quantification of (E) ubiquitin and (F) Ref(2)P in the heads of flies with and without nSMase knockdown in neurons using the elav-GAL4 driver. Results are normalized to Gba1b mutants without RNAi expression. At least 3 independent experiments were performed. Error bars represent SEM. *p < 0.05 by Student t-test.