Journal: Investigative ophthalmology & visual science

Article Title: ER Stress and Mitochondrial Perturbations Regulate Cell Death in Retinal Detachment: Exploring the Role of HIF1α.

doi: 10.1167/iovs.65.11.39

Figure Lengend Snippet: FIGURE 5. Hypoxia and retinal detachment activate mitophagy markers in retinal cells. (A) Representative immunoblots of PINK1, Parkin and FUNDC1 proteins from 661W cells subjected to 8 hours, 18 hours, and 24 hours of hypoxia. Blots are representative of three independent experiments in each condition; α-tubulin and GAPDH were used as loading controls. (B) Representative immunocytochemistry images of LC3B (green) and FUNDC1 (red) stained 661W cells subjected to 24 hours of hypoxia compared to normoxic control. Scale bar: 64 μm. (C) Representative immunoblots of Parkin and FUNDC1 proteins from retinas of C57BL/6J mice 1 day and 3 days following retinal detachment. Blots are representative of three independent animals in each condition; α-tubulin was used as the loading control. (D) Relative mRNA expression of Pink1 obtained at 1 dprd and 3 dprd compared to the contralateral eye (fellow). Pum-1 was used as a housekeeping control gene for normalization. (E) Relative mRNA expression of Prkn obtained at 1 dprd and 3 dprd compared to the contralateral eye (fellow). Pum-1 was used as a housekeeping control gene for normalization. Bar graphs represent mean ± SEM. Statistical analysis was performed using one-way ANOVA with repeated measures followed by Tukey’s test. *P < 0.05; **P < 0.01.

Article Snippet: The following antibodies were used: antimouse CHOP (#2895, 1:1000; Cell Signaling Technology, Danvers, MA, USA); anti-rabbit pIRE1α (#NB100-2323SS, 1:1000; Novus Biologicals, Littleton, CO, USA); anti-mouse BiP/Grp78 (#610979, 1:1000; BD Biosciences, Franklin Lakes, NJ, USA); anti-mouse mono-and polyubiquitinylated conjugates (#BML-PW8810, 1:1000; Enzo Life Sciences, Long Island, NY, USA); anti-mouse caspase 3 (#NB100-56113, 1:1000; Novus Biologicals); anti-mouse HIF1α (#MAB1536, 1:1000; R&D Systems, Minneapolis, MN, USA), anti-rabbit FUNDC1 (#NBP1-81063, 1:1000; Novus Biologicals); antirabbit PINK1 (#NB100-493, 1:1000; Novus Biologicals); antirabbit Parkin (#NBP2-67017, 1:1000; Novus Biologicals); and anti-mouse PGC-1α (#sc-517380, 1:500; Santa Cruz Biotechnology, Dallas, TX, USA).

Techniques: Western Blot, Immunocytochemistry, Staining, Control, Expressing

Journal: Investigative ophthalmology & visual science

Article Title: ER Stress and Mitochondrial Perturbations Regulate Cell Death in Retinal Detachment: Exploring the Role of HIF1α.

doi: 10.1167/iovs.65.11.39

Figure Lengend Snippet: FIGURE 6. HIF1α is associated with mitophagy induction after RD in retinal cells. (A) Representative immunoblots of HIF1α protein from 661W cells subjected to 8 hours, 18 hours, and 24 hours of hypoxia. Blots are representative of three independent experiments in each condition; α-tubulin was used as the loading control. (B) Representative immunoblots of HIF1α protein from retinas of C57BL/6J mice at 1 dprd and 3 dprd. Blots are representative of three independent animals in each condition; α-tubulin was used as the loading control. (C) Relative mRNA expression of Hspa5 obtained from retinas of Rho/Cre+ and HIF1αrod mice at 3 dprd compared to the contralateral eye (fellow). Pum-1 was used as a housekeeping control gene for normalization. (D) Relative mRNA expression of Ddit3 obtained from retinas of Rho/Cre+ and HIF1αrod mice at 3 dprd compared to the contralateral eye (fellow). Pum-1 was used as a housekeeping control gene for normalization. (E) Relative mRNA expression of Pgc1a and Tfam obtained from retinas of Rho/Cre+ and HIF1αrod mice at 3 dprd compared to the contralateral eye (fellow). Pum-1 was used as a housekeeping control gene for normalization. (F) Representative immunoblots of Parkin and FUNDC1 proteins from retinas of Rho/Cre+ and HIF1αrod mice at 3 dprd compared to the contralateral eye (fellow). GAPDH and α-tubulin were used as the loading control. (G) Relative mRNA expression of Prkn obtained from retinas of Rho/Cre+ and HIF1αrod mice at 3 dprd compared to the contralateral eye (fellow). Pum-1 was used as a housekeeping control gene for normalization. (H) Relative mRNA expression of Pink obtained from retinas of Rho/Cre+ and HIF1αrod mice at 3 dprd compared to the contralateral eye (fellow). Pum-1 was used as a housekeeping control gene for normalization. Bar graphs represent mean ± SEM. Statistical analysis was performed using two-way ANOVA with repeated measures followed by Tukey’s test. *P < 0.05; **P < 0.01; ***P < 0.001.

Article Snippet: The following antibodies were used: antimouse CHOP (#2895, 1:1000; Cell Signaling Technology, Danvers, MA, USA); anti-rabbit pIRE1α (#NB100-2323SS, 1:1000; Novus Biologicals, Littleton, CO, USA); anti-mouse BiP/Grp78 (#610979, 1:1000; BD Biosciences, Franklin Lakes, NJ, USA); anti-mouse mono-and polyubiquitinylated conjugates (#BML-PW8810, 1:1000; Enzo Life Sciences, Long Island, NY, USA); anti-mouse caspase 3 (#NB100-56113, 1:1000; Novus Biologicals); anti-mouse HIF1α (#MAB1536, 1:1000; R&D Systems, Minneapolis, MN, USA), anti-rabbit FUNDC1 (#NBP1-81063, 1:1000; Novus Biologicals); antirabbit PINK1 (#NB100-493, 1:1000; Novus Biologicals); antirabbit Parkin (#NBP2-67017, 1:1000; Novus Biologicals); and anti-mouse PGC-1α (#sc-517380, 1:500; Santa Cruz Biotechnology, Dallas, TX, USA).

Techniques: Western Blot, Control, Expressing

Journal: Investigative Ophthalmology & Visual Science

Article Title: ER Stress and Mitochondrial Perturbations Regulate Cell Death in Retinal Detachment: Exploring the Role of HIF1α

doi: 10.1167/iovs.65.11.39

Figure Lengend Snippet: Hypoxia and retinal detachment activate mitophagy markers in retinal cells. ( A ) Representative immunoblots of PINK1, Parkin and FUNDC1 proteins from 661W cells subjected to 8 hours, 18 hours, and 24 hours of hypoxia. Blots are representative of three independent experiments in each condition; α-tubulin and GAPDH were used as loading controls. ( B ) Representative immunocytochemistry images of LC3B ( green ) and FUNDC1 ( red ) stained 661W cells subjected to 24 hours of hypoxia compared to normoxic control. Scale bar : 64 µm. ( C ) Representative immunoblots of Parkin and FUNDC1 proteins from retinas of C57BL/6J mice 1 day and 3 days following retinal detachment. Blots are representative of three independent animals in each condition; α-tubulin was used as the loading control. ( D ) Relative mRNA expression of Pink1 obtained at 1 dprd and 3 dprd compared to the contralateral eye (fellow). Pum-1 was used as a housekeeping control gene for normalization. ( E ) Relative mRNA expression of Prkn obtained at 1 dprd and 3 dprd compared to the contralateral eye (fellow). Pum-1 was used as a housekeeping control gene for normalization. Bar graphs represent mean ± SEM. Statistical analysis was performed using one-way ANOVA with repeated measures followed by Tukey's test. * P < 0.05; ** P < 0.01.

Article Snippet: After fixation, the cells were permeabilized with 0.1% Triton X-100, blocked using 2% BSA, and subsequently subjected to an overnight incubation with the antibodies against FUNDC1 (1:250; Novus Biologicals) or anti-mouse LC3B (#83506, 1:250; Cell Signaling Technology).

Techniques: Western Blot, Immunocytochemistry, Staining, Control, Expressing

Journal: Investigative Ophthalmology & Visual Science

Article Title: ER Stress and Mitochondrial Perturbations Regulate Cell Death in Retinal Detachment: Exploring the Role of HIF1α

doi: 10.1167/iovs.65.11.39

Figure Lengend Snippet: HIF1α is associated with mitophagy induction after RD in retinal cells. ( A ) Representative immunoblots of HIF1α protein from 661W cells subjected to 8 hours, 18 hours, and 24 hours of hypoxia. Blots are representative of three independent experiments in each condition; α-tubulin was used as the loading control. ( B ) Representative immunoblots of HIF1α protein from retinas of C57BL/6J mice at 1 dprd and 3 dprd. Blots are representative of three independent animals in each condition; α-tubulin was used as the loading control. ( C ) Relative mRNA expression of Hspa5 obtained from retinas of Rho/Cre + and HIF1αΔrod mice at 3 dprd compared to the contralateral eye (fellow). Pum-1 was used as a housekeeping control gene for normalization. ( D ) Relative mRNA expression of Ddit3 obtained from retinas of Rho/Cre + and HIF1αΔrod mice at 3 dprd compared to the contralateral eye (fellow). Pum-1 was used as a housekeeping control gene for normalization. ( E ) Relative mRNA expression of Pgc1a and Tfam obtained from retinas of Rho/Cre + and HIF1αΔrod mice at 3 dprd compared to the contralateral eye (fellow). Pum-1 was used as a housekeeping control gene for normalization. ( F ) Representative immunoblots of Parkin and FUNDC1 proteins from retinas of Rho/Cre + and HIF1αΔrod mice at 3 dprd compared to the contralateral eye (fellow). GAPDH and α-tubulin were used as the loading control. ( G ) Relative mRNA expression of Prkn obtained from retinas of Rho/Cre+ and HIF1αΔrod mice at 3 dprd compared to the contralateral eye (fellow). Pum-1 was used as a housekeeping control gene for normalization. ( H ) Relative mRNA expression of Pink obtained from retinas of Rho/Cre + and HIF1αΔrod mice at 3 dprd compared to the contralateral eye (fellow). Pum-1 was used as a housekeeping control gene for normalization. Bar graphs represent mean ± SEM. Statistical analysis was performed using two-way ANOVA with repeated measures followed by Tukey's test. * P < 0.05; ** P < 0.01; *** P < 0.001.

Article Snippet: After fixation, the cells were permeabilized with 0.1% Triton X-100, blocked using 2% BSA, and subsequently subjected to an overnight incubation with the antibodies against FUNDC1 (1:250; Novus Biologicals) or anti-mouse LC3B (#83506, 1:250; Cell Signaling Technology).

Techniques: Western Blot, Control, Expressing

Journal: Science signaling

Article Title: The mitophagy effector FUNDC1 controls mitochondrial reprogramming and cellular plasticity in cancer cells

doi: 10.1126/scisignal.aaz8240

Figure Lengend Snippet: (A) PC3 cells transfected with control non-targeting siRNA (siCtrl) or FUNDC1-directed pooled siRNA (siFND1) were labeled with Talin-RFP and analyzed for focal adhesion (FA) dynamics by time-lapse videomicroscopy. Representative images at 0 h and 2 h from 3 independent experiments are shown. (B) The rate (events/h) of FA assembly (top) or disassembly (bottom) was quantified from the cells in (A). Each point corresponds to FA events in an individual cell (11–14 cells per condition). Mean±SD (N=3 independent experiments per group). *, p=0.02–0.04. (C and D) PC3 cells transfected with siCtrl or siFND1 were analyzed for cell motility in 2D contour plots by time-lapse video microscopy (C) with quantification of speed of cell movements (D, top) and distance traveled by individual cells (D, bottom). Each tracing in (C) corresponds to the movements of an individual cell (49 cells per condition) in a representative experiment. The cutoff velocities for slow (blue, <0.6 μm/min)- or fast (orange, >0.6 μm/min)-moving cells are indicated. Mean±SD (N=3 independent experiments per group). ***, p<0.0001. (E) PC3 cells transfected with siCtrl or siFND1 were analyzed for 2D chemotaxis in a rose plot (top) with quantification of the forward migration index (bottom). Arrows indicate the direction of the chemotactic gradient. Each point corresponds to an individual cell (89 cells per condition; N=3 independent experiments per group). (F) PC3 cells transfected with siCtrl or siFND1 were analyzed for invasion across Matrigel-coated Transwell inserts. Each point corresponds to an individual determination. Mean±SD (N=3 independent experiments per group). ***, p=0.0001. (G) The indicated prostate cancer cell lines were transfected with vector or FND1 cDNA and analyzed for Matrigel invasion. Mean±SD (N=3 independent experiments per group). **, p=0.001; ***, p <0.0001 – 0.0005. (H and I) PC3 cells transfected with vector or FND1 cDNA were analyzed by Western blotting (H, representative blot of three independent experiments) and the intensity of phosphorylated (p) FAK protein band was quantified by densitometry (I). Mean±SD (N=3 independent experiments per group). ***, p=0.0008. (J) PC3 cells transfected with siCtrl or siFND1 were analyzed for Matrigel invasion in the presence or absence of FAK-directed siRNA (siFAK). Mean±SD (N=3 independent experiments per group). ***, p<0.0001.

Article Snippet: Antibodies to FUNDC1 (Aviva System Biology, diluted 1:500), Drp1 (clone D6C7, Cell Signaling cat. #8570 diluted 1:1000), Ser 616 -phosphorylated Drp1 (Cell Signaling cat. #3455 diluted 1:1000), Drp1 (Cell Signaling cat. #5391 diluted 1:1000), Tyr 925 -phosphorylated FAK (Cell Signaling cat. #3284 diluted 1:1000), FAK (Cell Signaling cat. n. #3285), FLAG (clone M-2, Sigma cat. #F1804 diluted 1:5000) and β-actin (clone AC-15, Sigma cat. #A5441 diluted 1:100000) were used for Western blotting.

Techniques: Transfection, Labeling, Microscopy, Chemotaxis Assay, Migration, Plasmid Preparation, Western Blot

Journal: Science signaling

Article Title: The mitophagy effector FUNDC1 controls mitochondrial reprogramming and cellular plasticity in cancer cells

doi: 10.1126/scisignal.aaz8240

Figure Lengend Snippet: (A) PC3 cells were fractionated in submitochondrial compartments and analyzed by Western blotting. MTE, mitochondrial extracts; OM, outer membrane; IM, inner membrane; IMS, intermembrane space; Mat, matrix. Data are representative of 2 independent experiments. (B) Ingenuity pathway analysis of a mitochondrial FUNDC1 interactome identified by LC-MS/MS proteomics (N=1 independent experiment). The fold-induction for FUNDC1-associated proteins compared to Flag-vector are indicated with red color intensity. (C) PC3 cells transfected with Flag-vector or Flag-FND1 cDNA were immunoprecipitated (IP) with an antibody to Flag and immune complexes were analyzed by Western blotting. Data are representative of 2 independent experiments. WCE, whole cell extracts. (D) PC3 cells were immunoprecipitated with non-binding IgG or an antibody to endogenous ATP5C1 and immune complexes were analyzed by Western blotting. Data are representative of 2 independent experiments. WCE, whole cell extracts. (E) PC3 cells transfected with siCtrl or siFND1 were reconstituted with FND1 cDNA and analyzed for complex V (C.V) activity. Tracings from two independent experiments (Exp) are shown. Mean±SD of 3 technical replicates per experiment. (F) Citrate synthase-normalized complex V (C.V) activity as in (E) was quantified. Two independent experiments (Exp) are shown. Mean±SD of 3 technical replicates per experiment. (G) PC3 cells stably transduced with pLKO or shFND1 were reconstituted with Flag-LonP1 cDNA, and Flag-eluted LonP1 immune complexes (Elu) (top, representative blot of 3 independent experiments) were analyzed for LonP1 proteolytic activity (bottom). TCE, total cell extracts. Mean±SD (N=3 independent experiments per group). (H and I) PC3 cells transfected with siCtrl or siFND1 were reconstituted with LonP1 cDNA, extracted at increasing concentrations of NP-40 and detergent-insoluble protein bands corresponding to complex V subunits, ATP5C1, ATP5O and ATP5B were analyzed by Western blotting (H, representative blot of 2 independent experiments) and quantified by densitometry (I). VDAC was a control. Two independent experiments (Exp) are shown (I).

Article Snippet: Antibodies to FUNDC1 (Aviva System Biology, diluted 1:500), Drp1 (clone D6C7, Cell Signaling cat. #8570 diluted 1:1000), Ser 616 -phosphorylated Drp1 (Cell Signaling cat. #3455 diluted 1:1000), Drp1 (Cell Signaling cat. #5391 diluted 1:1000), Tyr 925 -phosphorylated FAK (Cell Signaling cat. #3284 diluted 1:1000), FAK (Cell Signaling cat. n. #3285), FLAG (clone M-2, Sigma cat. #F1804 diluted 1:5000) and β-actin (clone AC-15, Sigma cat. #A5441 diluted 1:100000) were used for Western blotting.

Techniques: Western Blot, Liquid Chromatography with Mass Spectroscopy, Plasmid Preparation, Transfection, Immunoprecipitation, Binding Assay, Activity Assay, Stable Transfection, Transduction

Journal: Science signaling

Article Title: The mitophagy effector FUNDC1 controls mitochondrial reprogramming and cellular plasticity in cancer cells

doi: 10.1126/scisignal.aaz8240

Figure Lengend Snippet: (A and B) PC3 (top), C42B (middle) or DU145 (bottom) cells were transfected with vector or FND1 cDNA and analyzed for Matrigel invasion (A) or cell proliferation by direct cell counting (B). Mean±SD (N=3 or 5 independent experiments per condition). **, p=0.001; ***, p <0.0001–0.0005. (C and D) PC3 cells stably transduced with pLKO or FUNDC1-directed shRNA (shFND1) were analyzed for colony formation (C) and quantified after 14 d (D). Mean±SD (N=3 independent experiments per group). ***, p<0.0001. (E) PC3 cells transduced with pLKO or shFND1 were engrafted subcutaneously onto the flanks of immunocompromised mice (N=10 animals/group) and tumor growth was quantified at the indicated time intervals. Each line corresponds to an individual tumor. The mean±SD tumor size (mm3) for each animal group harvested at day 27 is indicated, p<0.0001. (F) Representative tumor samples from each animal group in (E) were excised at the end of the experiment and analyzed for Ki67 staining by immunohistochemistry. Representative images are shown. Scale bar, 100 μm. (G and H) Representative lung samples isolated from each animal group in (E) were stained with an antibody to human mitochondria by immunohistochemistry (G, representative images) and the number of lung metastatic foci was quantified (H). Scale bar, 100 μm. Mean±SD (N=25 determinations per group). ***, p<0.0001. (I) PC3 cells stably transduced with pLKO or shFND1 were injected into the spleen of immunocompromised mice (N=5 animals per group) and metastatic foci to the liver were quantified after 11 days by histology. Mean±SD (N=8–10 determinations per group). ***, p=0.0009. (J and K) RNA-seq expression data from 33 cancer types (TCGA) for genes (Spearman r>0.2; p<0.05) positively (J, GO:0046034, ATP metabolism) or negatively (K, GO:0000302, response to ROS; GO:0048870, cell motility; GO:0008283, cell proliferation) correlated with FUNDC1 (FND1) expression.

Article Snippet: Antibodies to FUNDC1 (Aviva System Biology, diluted 1:500), Drp1 (clone D6C7, Cell Signaling cat. #8570 diluted 1:1000), Ser 616 -phosphorylated Drp1 (Cell Signaling cat. #3455 diluted 1:1000), Drp1 (Cell Signaling cat. #5391 diluted 1:1000), Tyr 925 -phosphorylated FAK (Cell Signaling cat. #3284 diluted 1:1000), FAK (Cell Signaling cat. n. #3285), FLAG (clone M-2, Sigma cat. #F1804 diluted 1:5000) and β-actin (clone AC-15, Sigma cat. #A5441 diluted 1:100000) were used for Western blotting.

Techniques: Transfection, Plasmid Preparation, Cell Counting, Stable Transfection, Transduction, shRNA, Staining, Immunohistochemistry, Isolation, Injection, RNA Sequencing Assay, Expressing

Journal: Science signaling

Article Title: The mitophagy effector FUNDC1 controls mitochondrial reprogramming and cellular plasticity in cancer cells

doi: 10.1126/scisignal.aaz8240

Figure Lengend Snippet: (A) Heatmap of changes in metabolite levels in PC3 cells transfected with siCtrl or siFND1. Fold changes, p values and false discovery rate (FDR) are indicated. Data are from a representative experiment (siCtrl) or two independent experiments in triplicate (siFND1, 1A-C; 2A-C). (B) Schematic diagram of mitochondrial bioenergetics and ROS pathways affected by FUNDC1 silencing as in (A). (C) PC3 cells transfected with siCtrl or siFND1were analyzed for oxygen consumption rates (OCR) on a Seahorse XFe96 Bioenergetics Flux Analyzer. Tracings from two independent experiments (Exp) are shown. Mean±SD of 3 technical replicates per experiment. (D) PC3 cells transfected with siCtrl or siFND1were analyzed for MitoSox reactivity and mitochondrial membrane potential (TMRE) by flow cytometry. Mean±SD (N=3–4 independent experiments per group). **, p=0.001. (E) PC3 cells transfected with siCtrl or siFND1were analyzed for MitoSox reactivity in the presence or absence of the superoxide scavenger, MnTBAP, by flow cytometry. Mean±SD (N=3 independent experiments per group). **, p=0.005; *, p=0.03. (F) PC3 cells transfected with siCtrl or siFND1were analyzed for speed of mitochondrial movements (top) or distance traveled by individual mitochondria (bottom) with or without MnTBAP. Each symbol corresponds to the tracked movement of an individual mitochondrion. Data are representative of 3 independent experiments. The mean±SD of mitochondrial speed (siCtrl compared to siFND1, p<0.0001; siFND1 compared to siFND1+MnTBAP, p=0.002) and distance traveled (siCtrl compared to siFND1, p=0.001; siFND1 compared to siFND1+MnTBAP, p=0.0005) are indicated. (G) siCtrl- or siFND1-transfected PC3 cells were analyzed for cellular motility in 2D contour plots in the presence or absence of MnTBAP. Each tracing corresponds to the movements of an individual cell. The cut-off velocities for slow (<0.25 μm/min)- or fast (>0.25 μm/min)-moving cells are indicated. Data are representative of 3 independent experiments per group. The speed (μm/min) of cell motility (siCtrl compared to siFND1, p<0.0001; siFND1 compared to siFND1+MnTBAP, p=0.006) and distance (μm) traveled (siCtrl compared to siFND1, p<0.0001; siFND1 compared to siFND1+MnTBAP, p=0.006) are indicated. (H) PC3 cells transfected with siCtrl or siFND1 were analyzed for Matrigel invasion in the presence or absence of MnTBAP. Mean±SD (N=3 independent experiments per group). ***, p<0.0001. (I) PC3 cells transfected with siCtrl or siFND1 were analyzed for cell proliferation in the presence or absence of MnTBAP by direct cell counting. Mean±SD (N=4 independent experiments per group). **, p=0.03.

Article Snippet: Antibodies to FUNDC1 (Aviva System Biology, diluted 1:500), Drp1 (clone D6C7, Cell Signaling cat. #8570 diluted 1:1000), Ser 616 -phosphorylated Drp1 (Cell Signaling cat. #3455 diluted 1:1000), Drp1 (Cell Signaling cat. #5391 diluted 1:1000), Tyr 925 -phosphorylated FAK (Cell Signaling cat. #3284 diluted 1:1000), FAK (Cell Signaling cat. n. #3285), FLAG (clone M-2, Sigma cat. #F1804 diluted 1:5000) and β-actin (clone AC-15, Sigma cat. #A5441 diluted 1:100000) were used for Western blotting.

Techniques: Transfection, Flow Cytometry, Cell Counting

Journal: bioRxiv

Article Title: MTCH2 is a mitochondrial outer membrane protein insertase

doi: 10.1101/2022.09.15.508165

Figure Lengend Snippet: (A) As in for a set of outer membrane reporters including a signal-anchored protein (USP30), a mitochondrial TA (MAVS) and a multipass protein (FUNDC1). (B) As in (A) but in either wild-type or MTCH2 KO K562 CRISPRi cells expressing guides targeting MTCH1 for knock-down. (C) Volcano plot of the genome-wide CRISPRi screen with MTCH1 and MTCH2 highlighted.

Article Snippet: The following antibodies were used in this study: MTCH2 (ab113707, Abcam, UK); FUNDC1 (OAAB12808, Aviva systems biology, USA); CYB5B (HPA007893, Atlas antibodies, USA); MIRO2 (RHOT2) (ab224089, Abcam, UK); CYC1 (4272, Cell signaling technology, USA); SYNJ2BP (OMP25) (15666-1-AP, Proteintech, USA); EMC3 (67205, Proteintech, USA); VDAC1 (sc-390996, Santa Cruz Biotech, USA); mitofilin (ab110329, Abcam, UK); SAMM50 (ab133709, Abcam, UK); ATP13A1 (16244-1-AP, Proteintech, USA); tubulin (T9026, Sigma-Aldrich, USA).

Techniques: Membrane, Expressing, Knockdown, Genome Wide