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. 2018 Feb 5;217(2):635–647. doi: 10.1083/jcb.201606095

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© 2018 Gong et al.

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Figure 7. — HSP90 contributes to the cytoprotective effects of HN. (A) CoIP of HN and HSP90 using GFP antibody in GFP-only or GFP-HN–overexpressing H9C2 cells (left) and HSP90 antibody in mouse liver lysate (right). IP, immunoprecipitation. (B) Colocalization of LysoTracker, HSP90, and HN in H9C2 cells. Bars: (main images) 10 µm; (magnifications) 2 µm. Arrowheads show colocalization. (C) L2A stability in isolated rat liver lysosomes incubated in an isotonic buffer in the presence or absence of HNG. Top: Representative immunoblots (IBs). Bottom: Quantification. n = 2. (D) Immunoblot and quantification of substrate uptake in the lysosomes isolated from rat liver with the presence of geldanamycin (Gelda), HNG, and HNG + geldamycin. (E) H9C2 cell viability at the absence and presence of HNG and 17-AAG with or without 1 mM PQ. Colocalization was analyzed in both Pearson’s correlation coefficient (Rr) and Manders’s Overlap coefficient (R). Differences with control were significant for *, P < 0.05; **, P < 0.01. Error bars show SEM.