Fig 2: MA-5 regulated the Bnip3-related mitophagy via the MAPK–ERK–Yap pathways. a through c Western blotting was used to analyze the changes in Yap and Bnip3. siRNA was used to silence Yap expression (si-YAP), which also reduced the expression of Bnip3. d The immunofluorescence assay of mitophagy via mitochondria and lysosome co-staining. The Yap siRNA also reduced the mitophagy. e through g SCH772984 was used to inhibit ERK activity. The ERK inhibitor also reduced Yap expression. *p < 0.05 vs. control (Ctrl) group; #p < 0.05 vs. TNFα group; †p < 0.05 vs. TNFα+MA-5 group

Fig 3: MFD increases lipid oxidation in the lungs during (a) acute and (b) chronic Mtb infection and protects from necrosis during chronic infection. Immunoblot analysis of adipogenesis marker (adiponectin), lipid oxidation marker (p-Perilipin, ATGL, p-HSL, and PPARα), and necrosis marker (BNIP3) in the lung lysates of RD-fed and MFD-fed uninfected and infected C57BL/6 mice during acute infection (30 DPI) and chronic (3 months) post-infection. The error bars represent standard error of the mean. * p < 0.05, ** p < 0.01, and *** p < 0.001 between indicated groups.

Fig 4: DUSP1 overexpression was involved in mitophagy inhibition via Bnip3. A-F. Western blots was used to analyze the proteins related to mitophagy. The siRNA against Bnip3 was used to knockdown the expression of Bnip3, which was considered as the negative control group. G. The immunofluorescence assay of mitochondria and lysosome. In the amplified panel, the yellow fluorescence is indicative of mitophagy. H. The ATP production was measured with DUSP1 overepxression or Bnip3 silence. I-M. The OCR assay was used to obverse the mitochondrial respiratory function. Data are shown as the means ± SEM. O-N. TUNEL assay was used to explore the role of Bnip3-related mitophagy in cell death. #P < 0.05 vs the control group; *P < 0.05 vs HR+Ad-ctrl group.

Fig 5: The expression of BNIP3 in UM tissues is detected by immunohistochemistry staining. (A) Representative image of the low expression of BNIP3. The score calculated by ImageJ-pro is 44.81. Scale bar: 50 um. (B) Representative image of the high expression of BNIP3. The score calculated by ImageJ-pro is 89.27.