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. 2015 Jul 13;4:e07366. doi: 10.7554/eLife.07366

Figure 6. Disruption of the tubular auto-lysosomal network correlates with increased poly-Ubiquitin aggregates, impaired mitochondria and increased lipofuscin granules.

(A, B) Wild type (A) and VCP-RNAi (B) expressing muscles were fixed and stained with a poly-Ubiquitin antibody. Nuclei with localized poly-Ubiquitin staining are apparent in A. Nuclei are indicated (dashed circle) in B. (C, D) Wild type animals were treated with DMSO (C) or the VCP-specific inhibitor DBeQ (D), fixed and stained with a poly-Ubiquitin antibody. (E) Quantitation of the number of poly-Ubiquitin aggregates per 50 µm2 from wild type muscles treated with DMSO for 4 hr or DBeQ for various times (n = 9, *p < 0.05, **p < 0.01). (F) Localization of Spin-GFP and poly-Ubiquitin in muscles expressing VCP-RNAi. (G, H) Mitotracker-C2TMRos staining in control (G) and VCP-RNAi (H) muscles. (I–K) Autofluoresence at 488 nm and lysotracker staining in wild type (I), muscles expressing VCP-RNAi (J), and wild type muscles treated with the VCP-specific inhibitor DBeQ for 4 hr (K).

DOI: http://dx.doi.org/10.7554/eLife.07366.015

Figure 6.

Figure 6—figure supplement 1. Loss of tubular lysosomes correlates with impaired muscle function.

Figure 6—figure supplement 1.

(A) Wild type and VCP-RNAi gross muscle morphology. (B) BG57-Gal4 control animals and animals expressing VCP-RNAi in muscles were assessed for their crawling ability on a petri dish. (C) The total distance traveled in 1 min was measured for each animal and averaged (n = 7, **p < 0.01). (C) Representative traces for wildtype muscles and muscles expressing VCP-RNAi. (D) Quantitation of EPSPs, mEPSPs and resistance input (n = 10, *p < 0.05, **p < 0.01).
DOI: http://dx.doi.org/10.7554/eLife.07366.016
Figure 6—figure supplement 2. Lysosomal acidity and Cathepsin processing are maintained in VCP-RNAi expressing muscles.

Figure 6—figure supplement 2.

(A) Spin-GFP co-imaging with lysotracker in muscles expressing VCP-RNAi. (B) Western blot analysis of Cathepsin L processing in the genotypes indicated. Tubulin serves as a loading control.
DOI: http://dx.doi.org/10.7554/eLife.07366.017