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. 2014 Feb 10;111(8):3116–3121. doi: 10.1073/pnas.1318951111

Fig. 4.

Fig. 4.

Parkin-mediated mitophagy maintains insulin secretion under diabetic conditions. (A) Construction of the stable MIN6 β-cell lines overexpressing Parkin. Representative immunoblots are shown from three independent experiments. (B) Excitation ratio (590/430 nm) of pMT-mKeima-Red–overexpressing stable cell lines incubated with HG and PA. Baf-A1–treated cells were used as a negative control for the induction of mitophagy. The experiments were performed in eight replicates (n = 3). (C) The sequential state III OCR of isolated mitochondria from stable cell lines was assessed using the Seahorse XF24 analyzer. Representative results are shown, and the OCR of each complex was calculated from five replicates per sample (n = 3). (D) Glucokinase (GK) and hexokinase (HK) activities in stable cell lines. Results are shown from four independent experiments performed in duplicate. (E) ATP contents and (F and G) insulin secretion were measured by a luminometer and ELISA, respectively. The experiments were performed in duplicate (n = 4). Data are shown as the means ± SD. †P < 0.05 versus stable cell lines carrying control vectors under the same condition.