Figure 4

FASN regulates BLM-induced loss of mitochondrial membrane potential and mitochondrial dysfunction in AECs. FASN-overexpressing and knockdown cells treated with BLM (10 μg/mL) were stained with JC-1 for 30 min and analyzed by flow cytometry. (A) The proportion of depolarizing mitochondrial membrane potential (JC-1 green fluorescence) was increased in FASN knockdown cells, whereas it was decreased by FASN overexpression. Graph showing the ratio of the intensity of red to green fluorescence (JC-1 aggregate and JC-1 monomer) of flow cytometry. The data are expressed as the mean ± standard error of the mean (n = 4). *P < 0.05, vs Scramble or pCMV. #P < 0.05, vs Scramble + BLM or pCMV + BLM. (B) Protein expression of PINK1 was measured by Immunoblotting analysis in the lungs of FASN transgenic mice. The intensity of the PINK1 was quantified by densitometry, and the data were normalized to β-actin. Cropped images are displayed, uncropped blots are displayed in Supplementary Fig. 5. The data are expressed as the mean ± SEM. Error bars represent mean ± SEM (n = 6) *P < 0.05, vs. Doxy (−)/BLM (+). (C) Immunofluorescence analysis images of PINK1 (red) in AECIIs (SPC, green) of FASN transgenic mice along with the quantification of double positive cells (n = 6, each group). Scale bar, 50 µm. The data are expressed as the mean ± standard error of the mean. *P < 0.05, vs. Doxy (−)/BLM (−)panels × 400.