Fig 2: Possible mechanism(s) underlying in the mitophagic regulation of berberine in Sham- and TAC-operated mice treated with or without BBR. (A) Representative gel blots for each group by western blot (n = 3); (B) Mito-Parkin to VDAC ratio; (C) PINK1 to β-actin ratio; (D) p-Parkin(ser65) to β-actin ratio; (E) Total-Parkin to β-actin ratio; (F) FUNDC1 to β-actin ratio; (G) PHB2 to Vinculin ratio; (H) BNIP3L to Vinculin ratio. Mean ± SEM, * p < 0.05 vs. Sham-Veh group; # p < 0.05 vs. TAC-Veh group.

Fig 3: Empagliflozin sustains mitochondrial homeostasis in CMECs through the AMPKα1/ULK1/FUNDC1 pathway.In vivo, AMPKα1EKO, FUNDC1EKO or Tie2Cre (control) mice were assigned to the sham operation group or the myocardial I/R injury group. Empagliflozin (10 mg/kg/d) was administered seven days before myocardial I/R injury. In vitro, CMECs were isolated from I/R- or empagliflozin-treated hearts. The cells were cultured for 24 h and then used for functional analyses. (A–C) Immunofluorescence assays were used to assess the mitochondrial morphology in CMECs. The average mitochondrial length was recorded to reflect mitochondrial fission. At least 100 mitochondria from 10 CMECs were used to evaluate the number of CMECs with fragmented mitochondria. (D–F) Mitochondrial and cytoplasmic ROS levels in CMECs were determined using immunofluorescence assays. Mitochondrial ROS were assessed using MitoSOX™ Red, while cytoplasmic ROS were measured using CM-H2DCFDA. (G–I) The activity levels of intracellular anti-oxidative molecules such as GSH, SOD and GPX in CMECs were determined using commercially available ELISA kits. (J–K) JC-1 staining was used to determine the mitochondrial membrane potential in CMECs. A reduced red-to-green immunosignal of JC-1 indicates an abnormal mitochondrial membrane potential. Experiments were repeated at least three times and the data are shown as mean ± SEM (n = 6 mice or three independent cell isolations per group). *p < 0.05. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.

Fig 4: Fundc1 deletion attenuates IPC-mediated renoprotection in IRI kidneys. (A–B) Following IRI, levels of BUN and Cr were determined in various groups. (C–D) H&E staining for reperfused kidneys. Tubular injury index was determined. (E–F) TUNEL assay for IRI kidneys. The number of apoptotic cells was evaluated. (G–H) qPCR was used to observe pro-inflammation factors. Experiments were repeated at least three times and data are shown as mean ± SEM (n = 6 mice per group). Fundc1f/f mice in sham group were used as the normalizer for all the conditions. *P < 0.05.

Fig 5: Hypoxia-inducible factor-BNIP3/FUNDC1-mediated mitophagy prevented bevacizumab-induced ROS accumulation and ROS-induced apoptosis in OIR mouse retina: (A–C) OIR mice received intravitreal injection of bevacizumab alone or bevacizumab plus LW6, Representative immunofluorescence images showing LC3 co-staining with HIF, BNIP3, or FUNDC1 in retina of OIR model. (D) OIR mice received intravitreal injection of bevacizumab alone or bevacizumab plus CQ injection, Representative TUNEL staining imaging that CQ suppressed mitophagy and promotes apoptosis in bevacizumab-treated retina. Bar = 100 μm. (E) Representative fluorescence images showing that CQ interfered with mitophagy and promotes ROS production in bevacizumab-treated retina. Bar:100 μm. ns: no significance, *P < 0.05, **P < 0.01, ***P < 0.001.