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. 2020 Oct 23;38:101767. doi: 10.1016/j.redox.2020.101767

Fig. 4.

Fig. 4

Knockdown of Pink1 or Park2 inhibits OPTN recruitment to mitochondria during LPS treatment. (A) Co-localization of OPTN with mitochondria upon LPS treatment. RPTC cells were subjected to 24 h of 100 μg/ml LPS treatment. Mitochondria in these cells were then labeled with MitoTracker Red FM, while OPTN was stained by immunofluorescence. These cells were examined by confocal microscopy to show the co-localiztion of OPTN (green) and mitochondria (red) in LPS-treated cells (arrows). (B, C) Immunoblot analysis of OPTN accumulation in mitochondria and the effects of Pink1 or Park2 knockdown. RPTC cells transfected with siRNAs for Pink1 or Park2 or negative control siRNA (NC siRNA) were subjected to LPS treatment. These cells were harvested and fractionated to cytosolic (cyto) and mitochondrial (mito) fractions for immunoblot analysis of OPTN, COX4(mitochondrial marker), and GAPDH (cytosolic marker). (B) Representative blots. (C) Densitometry of OPTN. Mean ± SD. n = 3. *P < 0.001 vs. control group. #P < 0.01 vs. siRNA-LPS group. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)