MFN1 Antibody - #DF7543

FIGURE 1 Ezrin regulates mitochondrial fission and fusion and mtROS production in cells. Three Ezrin-specific siRNAs (the si-Ezr-001, si-Ezr-002 and si-Ezr-003) were transfected into HaCat cells for 48 h. (A) COIP was used to observe the specific binding of Ezrin, P-Ezrin (T567) and Drp1. (B) Immunofluorescence staining was used to observe the localization and expression of Ezrin and P-Ezrin (T567) in HaCat cells. (C) Western blot analysis was used to measure the expression of Ezrin, P-Ezrin (T567), Drp1, P-Drp1, Fis1, Mfn1, Mfn2 and Opa1. (D) Observation of mitochondrial morphology and mtROS by immunofluorescence staining of TOMM20 and MitoSOX. (E) ROS generation was observed after DCFH-DA staining. (F) Cellular immunofluorescence staining was used to observe the localization and expression of P-Drp1. (G) Mitotracker Red and Drp1 fluorescence staining were used to observe the translocation of Drp1 to mitochondria. *p 

Figure 3 JTE-013 inhibited the expression of mitochondrial fusion/fission protein and fibrosis marker proteins in lung tissues. (A): Immunofluorescence staining was used to detect the colocalization of Drp1 and Tom20 (scale bar = 200 μm). (B): TUNEL staining was used to detect apoptosis in lung tissue. (C): Western blot was used to detect the expression of Fis1, OPA1, and MFN1/2 and the phosphorylation and transformation of Drp1. (D): Immunohistochemical staining was used to detect the expression of α-SMA, COL1A1, and MMP-9 and their OD quantitative analysis chart (scale bar = 50 μm). (E): Western blot was used to detect the expression of α-SMA, COL1A1, and MMP-9 (** p < 0.01 vs. control group, ## p < 0.01 vs. BLM group, && p < 0.01 vs. BLM + JTE-013 group). Original Western blot available in Supplementary Materials.

Fig. 5 Effect of intervene in SIRT-1 and combine with OA on p62, Beclin-1, LC3B, SIRT-1, PGC-1a, MFN1, MFF and COX-IV protein expression in HepG2 cells. HepG2 cells were treated with 1.5 mM OA for 48 h, following T6 (2 μM) or CAY (20 μM) 2 h. a The p62, Beclin-1 and LC3B protein expression was detected by Western blotting. b The same method was used to detect the SIRT-1, PGC-1a, MFN1, MFF and COX-IV protein expression. The bands were normalized using GAPDH. The images were quantified with ImageJ. Data are presented as the mean ± SD; *P < 0.05, **P < 0.01 compared with CON group; #P < 0.05, ##P < 0.01 compared with OA group

Figure 3 Mitochondrial changes in activated microglia (M1). (a) Schematic diagram of intracellular mitochondrial fusion and fission process in microglia after OGD/R. (b) Western blot assay of mitochondrial fusion protein (Opa1 and Mfn1), TOM20, and cytochrome c in mitochondria of M0 and M1 microglia (n = 3). Results are displayed in a form of mean ± SD; ∗P < 0.05, ∗∗P < 0.01, and ∗∗∗P < 0.001. (c) Western blotting findings of mitochondrial fission protein including MFF, Fis1, Mid49, and Mid51 in mitochondria of M0 and M1 microglia (n = 3). Data presented are mean ± SD; ∗∗P < 0.01 and ∗∗∗P < 0.001. (d) Mitochondrial function in M1 microglia and M0 microglia was investigated by determining ATP (n = 3), mitochondria membrane potential (n = 3), and ROS (n = 3). The relative ratio of intracellular ATP was determined by calculating the ratio of level of ATP in M0-BV2 and M1-BV2 cells to level of ATP in M0-BV2 cells. Results are displayed in a form of mean ± SD. ∗∗P < 0.01 and ∗∗∗P < 0.001. (e) Morphology of intracellular mitochondria in BV2 cells visualized under TEM, scale bar: 1.0 μm.