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. 2013 Oct 1;123(11):4667–4680. doi: 10.1172/JCI67615

Figure 2. Impaired mitochondrial respiration in the hypothalamus is HSP60 dependent and induces oxidative stress.

Figure 2

(A) Western blot analysis of HSP60 in N25/2 cells infected with a lentiviral Hsp60 shRNA or scrambled shRNA. (B) Basal respiration and (C) maximal respiratory capacity measurements, displayed as area under the curve, of control and HSP60 KD cells. (D) Assessment of mitochondrial volume using MitoTracker green staining in control and HSP60 KD cells (n = 3 each). (E) Electron microscopy from mitochondria of control and HSP60 KD cells. Scope magnification, ×19,000; enlargement magnification, ×2.45; total magnification, ×46,550. (F) Mitochondrial DNA compared with genomic DNA content (n = 6 each). (G) Western blot analysis of nuclear- and mitochondrial-encoded genes in control (n = 2) and HSP60 KD cells n = 2 each). The experiment was performed twice with a total of four per group. (H) Gene expression analysis of analyzed nuclear- and mitochondrial-encoded proteins. (I) Citrate synthase activity in control and HSP60 KD cells (n = 8 each). (J) Quantification of superoxide accumulation using MitoSOX staining of control and HSP60 KD cells (n = 3 each). (K) Quantification of lipid peroxidation using TBARS assay on control (n = 5) and HSP60 KD cells (n = 6). Displayed values are the means ± SEM. *P ≤ 0.05; **P ≤ 0.01; ***P ≤ 0.001.