Journal: International Journal of Biological Sciences

Article Title: mtDNA-cGAS-STING axis-dependent NLRP3 inflammasome activation contributes to postoperative cognitive dysfunction induced by sevoflurane in mice

doi: 10.7150/ijbs.91543

Figure Lengend Snippet: Inhibiting mPTP-VDAC channel opening attenuates sevoflurane-induced mtDNA cytosolic escape and reduces cGAS-STING pathway activation in microglia. A BV2 cells were treated with 1 mM sevoflurane for 12 h after 1 µM CsA intervention for 30 min and 10 µM VBIT-4 intervention for 30 min. mtDNA levels of Nd1, Cytb, and D-loop in cytoplasm of BV2 cells were determined by real-time PCR ( n =3). B-I Protein levels of NLRP3, pro-IL-1β, IL-1β p17, cGAS, STING, p-TBK1 Ser172 , and p-IRF3 Ser396 in the BV2 cells were detected by western blot ( n =3). J-Q Protein levels of NLRP3, pro-IL-1β, IL-1β p17, cGAS, STING, p-TBK1 Ser172 , and p-IRF3 Ser396 in the BV2 cells were detected by western blot ( n =3). Data are expressed as the mean ± SD. Differences among multiple groups were performed using ANOVA. *P < 0.05, **P < 0.01, and ***P < 0.001.

Article Snippet: To evaluate the role of mitochondrial DNA in sevoflurane-induced inflammation in BV2 cells, we treated cells with the DRP1 inhibitor (Mdivi-1, 100 nM, MedChemExpress), the mPTP inhibitor (CsA, 1 µM, TargetMol, China) or the VDAC inhibitor (VBIT-4, 10 µM, TargetMol) 30 min before sevoflurane stimulation.

Techniques: Activation Assay, Real-time Polymerase Chain Reaction, Western Blot

Journal: International Journal of Biological Sciences

Article Title: mtDNA-cGAS-STING axis-dependent NLRP3 inflammasome activation contributes to postoperative cognitive dysfunction induced by sevoflurane in mice

doi: 10.7150/ijbs.91543

Figure Lengend Snippet: Schematic illustration. Sevoflurane promoted DRP1-dependent mitochondrial fission to release mtDNA into the cytoplasm via the mPTP-VDAC channel, which induced the cGAS-STING pathway-dependent NLRP3 inflammasome activation, resulting in neuroinflammation of microglia.

Article Snippet: To evaluate the role of mitochondrial DNA in sevoflurane-induced inflammation in BV2 cells, we treated cells with the DRP1 inhibitor (Mdivi-1, 100 nM, MedChemExpress), the mPTP inhibitor (CsA, 1 µM, TargetMol, China) or the VDAC inhibitor (VBIT-4, 10 µM, TargetMol) 30 min before sevoflurane stimulation.

Techniques: Activation Assay

Journal: Cell Reports Medicine

Article Title: Urolithin A improves muscle strength, exercise performance, and biomarkers of mitochondrial health in a randomized trial in middle-aged adults

doi: 10.1016/j.xcrm.2022.100633

Figure Lengend Snippet: Urolithin A confers a proteomic signature of improved mitochondrial metabolism and mitophagy in human skeletal muscle (A) Venn diagram summarizing pathway enrichment analysis results from proteomics data in vastus lateralis skeletal muscle. Data represent upregulated pathways with an adjusted p value <0.1 in subjects treated with placebo or with UA at 500 and 1,000 mg for 4 months compared with baseline. (B and C) Dot plots showing top enriched pathways (WikiPathways 2019 Human), ranked by protein ratio, in the UA 500- (B) and UA 1,000 mg (C) groups from (A). Dot color and size indicate adjusted p value and protein count, respectively. Significant pathways for the placebo group were filtered out to identify treatment-specific pathways. (D) Western blot analysis of protein lysates from vastus lateralis skeletal-muscle biopsies in subjects treated as above. For each subject, both baseline (Pre) and end-of-the-treatment (Post) samples were run, and membranes were probed for phospho-Parkin, total Parkin, and the mitochondrial proteins ATP5A (belonging to the OXPHOS complex V), UQCRC2 (complex IV), SDHB (complex II), and NDUFB8 (complex I). Tubulin and VDAC were included as markers of total and mitochondrial protein abundance, respectively. Dashed line separates samples from individual subjects. (n = 6 Pre and Post, biologically independent samples). (E) Quantification of phospho-Parkin over Tubulin protein intensity from western blots (WBs) in (D) (n = 6). Two-sided, paired t-test. (F) Quantification of NDUFB8 (left) and SDHB (right) protein intensity, normalized over VDAC from WBs in (D) (n = 6). ∗p < 0.05; ∗∗p < 0.01; two-sided, paired t test.

Article Snippet: The following primary antibodies were incubated overnight diluted in blocking buffer: UBE2N (SantaCruz, sc-376470, 1:3000), Tubulin (Proteintech, 10004185, 1:3000), phospho-Parkin S65 (Cell Signaling, #36866, 1:1000), Parkin (Cell Signaling, #4211, 1:1000), OXPHOS Antibody Cocktail (Abcam, ab110413, 1:2000), VDAC (Proteintech, 10866-1-AP, 1:1000).

Techniques: Western Blot, Quantitative Proteomics

Journal: Cell Reports Medicine

Article Title: Urolithin A improves muscle strength, exercise performance, and biomarkers of mitochondrial health in a randomized trial in middle-aged adults

doi: 10.1016/j.xcrm.2022.100633

Figure Lengend Snippet:

Article Snippet: The following primary antibodies were incubated overnight diluted in blocking buffer: UBE2N (SantaCruz, sc-376470, 1:3000), Tubulin (Proteintech, 10004185, 1:3000), phospho-Parkin S65 (Cell Signaling, #36866, 1:1000), Parkin (Cell Signaling, #4211, 1:1000), OXPHOS Antibody Cocktail (Abcam, ab110413, 1:2000), VDAC (Proteintech, 10866-1-AP, 1:1000).

Techniques: Recombinant, RNA Extraction, Software, Protease Inhibitor

Journal: The Journal of Biological Chemistry

Article Title: Plasma Membrane Origin of the Steroidogenic Pool of Cholesterol Used in Hormone-induced Acute Steroid Formation in Leydig Cells *

doi: 10.1074/jbc.M116.740928

Figure Lengend Snippet: Effect of inhibitors of the proteins associated with the steroidogenic metabolon on cholesterol trafficking from the plasma membrane in MA-10 cells transfected with mCherry-D4. A–D, time course of mCherry-D4 fluorescence associated with the plasma membrane in the presence of various inhibitors in the presence or absence of Bt2cAMP. A, aminoglutethimide (AMG), an inhibitor of CYP11A1. B, erastin, an inhibitor of VDAC. C, two concentrations of the START domain ligand, an inhibitor of the STAR protein. D, two concentrations of the CRAC domain ligand, an inhibitor of TSPO. E, progesterone production 2 h after exposure of the cells to various inhibitors. Data represent means ± S.D. of at least three independent experiments performed in triplicate; two-way ANOVA followed by Bonferroni's post hoc test (*) was used to calculate statistical significance; *, p < 0.05; **, ##, p < 0.01; ***, p < 0.001. CRAC, cholesterol recognition amino acid consensus sequence; CYP11A1, cytochrome P450 side chain cleavage enzyme; STAR, steroidogenic acute regulatory protein; TSPO, translocator protein; VDAC, voltage-dependent anion channel.

Article Snippet: The cell treatments were as follows: 1 m m Bt 2 cAMP (Sigma); 20 μ m 22 R -HC (Sigma); 20 μ m 22 S -HC (Sigma); 20 μ m progesterone; 20 μ m pregnenolone; 1 or 10 μ m TSPO CRAC domain ligand ( N -[2-(4-ethyl-5-[2-oxo-2-(4-toluidino)ethyl]sulfanyl-4 H -1,2,4-triazol-3-yl)ethyl]-4-methylbenzamide) ( 46 ); 100 μ m START domain of STAR protein ligand (16-[4-(difluoromethoxy)benzylidene]androst-5-ene-3,17-diol) ( 47 ); 0.67 m m dl -aminoglutethimide (Sigma), a CYP11A1 inhibitor, or 100 μ m erastin (Sigma), an inhibitor of VDAC.

Techniques: Transfection, Fluorescence, Sequencing

Journal: Cardiovascular research

Article Title: Mitochondrial pannexin1 controls cardiac sensitivity to ischaemia/reperfusion injury.

doi: 10.1093/cvr/cvad120

Figure Lengend Snippet: Figure 5 Panx1 modulates mitochondrial respiration. Western blots revealed lower Panx1 expression in B16-F10 (F10) cells as compared to isogenic B16-BL6 (BL6) cells (A). Representative traces of OCR measured by Seahorse in untreated B16-BL6 (black) and B16-F10 (grey) cells or after VDAC1 inhibition with 10 μM NSC15364 (B16-BL6: red; B16-F10: green) (B). Basal respiration was reduced in B16-F10 compared to B16-BL6 cells. N = 15, unpaired t-test. (C). Likewise, the maximal respiration rate was reduced in B16-F10 compared to B16-BL6 cells after VDAC1 inhibition with 10 μM of TRO19622 (N = 5, unpaired t-test) (D) or NSC15364 (N = 5, unpaired t-test) (E). Representative traces of calcium-induced mPTP opening in cardiac SSM from WT (black) and Panx1−/−

Article Snippet: Voltage-dependent anion channel 1 (VDAC1) inhibitors TRO19622 (Tocris) or NSC15463 (MedChemExpress) were used at a concentration of 10 μM, oligomycin at 20 μM, carbonyl cyanide 4−(trifluoromethoxy)phenylhydrazone (FCCP) at 1 μM and rotenone/antimycin at 5 μM.

Techniques: Western Blot, Expressing, Inhibition

Journal: Scientific Reports

Article Title: Hypothermia regulates mitophagy and apoptosis via PINK1/Parkin-VDAC 3 signaling pathway during oxygen-glucose deprivation/recovery injury

doi: 10.1038/s41598-025-89176-w

Figure Lengend Snippet: Effects of hypothermia on MMP and mPTP opening in model of OGD/R-induced BV2 mouse microglia cells damage. ( A ) The scatter diagram of MMP detected by flow cytometry. Under normal conditions, JC-1 aggregated in the mitochondrial matrix to multimer. When MMP was reduced, JC-1 could not aggregate in the mitochondrial matrix, but existed as a monomer. ( B - C ) The quantitative analysis of damaged MMP percentage. ( D , F ) The peak superposition diagram of mPTP opening tested by flow cytometry. Under normal conditions, the opening of mPTP was not excessively activated, and the fluorescence of BbcellProbe M61 was relatively higher. When mitochondria were damaged, the opening of mPTP was excessively activated, and the fluorescence of BbcellProbe M61 was relatively lower as well as the representative diagram shifts left compared with the base line. ( E , G ) The quantitative analysis of the mean fluorescence of BbcellProbe M61, which represented the opening of mPTP. ( H - J ) The western bands of VDAC3 and the quantitative analysis. ( K - L ) The mRNA expression of VDAC3 and the quantitative analysis. Data was presented as the mean ± SD ( n ≥ 3). * P < 0.05, vs. Sham group; # P < 0.05, vs. OGD group; & P < 0.05, vs. OGD/R-NormoT-2 h groups; $ P < 0.05, vs. OGD/R-NormoT-4 h group.

Article Snippet: The materials used in this study were as follows: microglial mouse cell lines (catalog # CL-0493, Procell); high glucose Dulbecco’s modified Eagle’s medium (DMEM; catalog # C12430500BT, Gibco); 10% fetal bovine serum (FBS; catalog # 26010074, Gibco); DMEM without glucose (catalog # 11966025, Gibco); modular incubator chamber (catalog # MTC101, billups-rothenberg); single flowmeter (catalog # SFM3001, billups-rothenberg); dulbecco’s phosphate-buffered Saline (D-PBS, catalog # Lvn1023, livning); 0.4% trypan blue stain (catalog # C0040 solarbio); electron microscope fixing solution, (catalog #G1102, Servicebio); JC-1 MMP Assay (catalog # HY-K0601, medchemexpress); BBcellProbe M61 mPTP detection kit (catalog # BB-48122, BestBio); RIPA Lysis Buffer (catalog # C-1053, Applygen); Phenylmethanesulfonyl fluoride (PMSF; catalog # A1100, Applygen); Protease inhibitor cocktail (catalog # P1265-1, Applygen); NcmBlot blocking buffer (catalog # P30500, Ncmbio); rabbit anti-PINK1 antibody (catalog # 23274-1-AP, proteintech); rabbit anti-Parkin antibody (catalog # 32833, cell signaling technology); mouse anti-Parkin antibody (catalog # 4211, cell signaling technology); rabbit anti-VDAC3 antibody (catalog # 55260-1-AP, proteintech); rabbit anti-Bcl2 antibody (catalog # 26593-1-AP, proteintech); rabbit anti- Beclin-1 antibody (catalog # 3495 S, cell signaling technology); rabbit anti-Beta Actin antibody (catalog # 20536-1-AP, proteintech); rabbit anti-GAPDH antibody (catalog # 10494-1-AP, proteintech); rabbit anti-Cleaved Caspase 3 antibody (catalog # 9664, cell signaling technology); rabbit anti-caspase3 antibody (catalog # 14220, cell signaling technology); rabbit anti- SQSTM1/p62 antibody (catalog # 39749, cell signaling technology); rabbit anti-Cytochrom C antibody (catalog # 10993-1-AP, proteintech); rabbit anti-LC3B antibody (catalog # 43566, cell signaling technology); goat anti-rabbit HRP-conjugated IgG secondary antibody (catalog # ZB-2301, zsbio); Goat anti-Mouse IgG (H + L) Secondary Antibody, DyLightTM 488 (catalog # 35502, Thermo fisher scientific); Goat anti-Rabbit IgG (H + L) Secondary Antibody, DyLightTM 594(catalog # 35560, Thermo fisher scientific); HiScript III RT SuperMix for qPCR (+ gDNA wiper), (catalog # R323-01, Vazyme Biotech); FastPure Complex Cell/Tissue Total RNA Isolation Kit (catalog # RC113-C1, Vazyme Biotech); Taq Pro Universal SYBR qPCR Master Mix (catalog # Q712-03, Vazyme Biotech); QuickBlockTM Blocking Buffer for Immunol Staining (catalog # P0260, Beyotime).

Techniques: Flow Cytometry, Fluorescence, Western Blot, Expressing

Journal: Scientific Reports

Article Title: Hypothermia regulates mitophagy and apoptosis via PINK1/Parkin-VDAC 3 signaling pathway during oxygen-glucose deprivation/recovery injury

doi: 10.1038/s41598-025-89176-w

Figure Lengend Snippet: Effects of hypothermia on Parkin-VDAC3 colocalization in model of OGD/R-induced BV2 mouse microglia cells damage. Subcellular localizations of Parkin (red), VDAC3 (green), and DAPI (blue) were observed. (Scale bars, 20 μm.) Colocalization analysis results: the 2D intensity histogram of colocalization and Pearson’s correlation coefficient (Pearson’s R value).

Article Snippet: The materials used in this study were as follows: microglial mouse cell lines (catalog # CL-0493, Procell); high glucose Dulbecco’s modified Eagle’s medium (DMEM; catalog # C12430500BT, Gibco); 10% fetal bovine serum (FBS; catalog # 26010074, Gibco); DMEM without glucose (catalog # 11966025, Gibco); modular incubator chamber (catalog # MTC101, billups-rothenberg); single flowmeter (catalog # SFM3001, billups-rothenberg); dulbecco’s phosphate-buffered Saline (D-PBS, catalog # Lvn1023, livning); 0.4% trypan blue stain (catalog # C0040 solarbio); electron microscope fixing solution, (catalog #G1102, Servicebio); JC-1 MMP Assay (catalog # HY-K0601, medchemexpress); BBcellProbe M61 mPTP detection kit (catalog # BB-48122, BestBio); RIPA Lysis Buffer (catalog # C-1053, Applygen); Phenylmethanesulfonyl fluoride (PMSF; catalog # A1100, Applygen); Protease inhibitor cocktail (catalog # P1265-1, Applygen); NcmBlot blocking buffer (catalog # P30500, Ncmbio); rabbit anti-PINK1 antibody (catalog # 23274-1-AP, proteintech); rabbit anti-Parkin antibody (catalog # 32833, cell signaling technology); mouse anti-Parkin antibody (catalog # 4211, cell signaling technology); rabbit anti-VDAC3 antibody (catalog # 55260-1-AP, proteintech); rabbit anti-Bcl2 antibody (catalog # 26593-1-AP, proteintech); rabbit anti- Beclin-1 antibody (catalog # 3495 S, cell signaling technology); rabbit anti-Beta Actin antibody (catalog # 20536-1-AP, proteintech); rabbit anti-GAPDH antibody (catalog # 10494-1-AP, proteintech); rabbit anti-Cleaved Caspase 3 antibody (catalog # 9664, cell signaling technology); rabbit anti-caspase3 antibody (catalog # 14220, cell signaling technology); rabbit anti- SQSTM1/p62 antibody (catalog # 39749, cell signaling technology); rabbit anti-Cytochrom C antibody (catalog # 10993-1-AP, proteintech); rabbit anti-LC3B antibody (catalog # 43566, cell signaling technology); goat anti-rabbit HRP-conjugated IgG secondary antibody (catalog # ZB-2301, zsbio); Goat anti-Mouse IgG (H + L) Secondary Antibody, DyLightTM 488 (catalog # 35502, Thermo fisher scientific); Goat anti-Rabbit IgG (H + L) Secondary Antibody, DyLightTM 594(catalog # 35560, Thermo fisher scientific); HiScript III RT SuperMix for qPCR (+ gDNA wiper), (catalog # R323-01, Vazyme Biotech); FastPure Complex Cell/Tissue Total RNA Isolation Kit (catalog # RC113-C1, Vazyme Biotech); Taq Pro Universal SYBR qPCR Master Mix (catalog # Q712-03, Vazyme Biotech); QuickBlockTM Blocking Buffer for Immunol Staining (catalog # P0260, Beyotime).

Techniques:

Journal: Scientific Reports

Article Title: Hypothermia regulates mitophagy and apoptosis via PINK1/Parkin-VDAC 3 signaling pathway during oxygen-glucose deprivation/recovery injury

doi: 10.1038/s41598-025-89176-w

Figure Lengend Snippet: Cellular signal pathway diagram of hypothermia at 2 h after temperature intervention. On the one hand, hypothermia promoted the expression of Cleaved PINK1 at first, and then recruited Parkin onto the outer mitochondrial membrane, thereby decreasing the expression of VDAC3 as well as SQSTM/p62, and then increased the ratio of LC3B II/LC3B I and reduced the release of Cyt C via decreasing mPTP opening, subsequently reduced the expression of downstream proapoptotic proteins such as cleaved caspase3 and caspase3 during OGD/R. Finally, hypothermia provided neuroprotective effects via alleviating apoptosis and promoting mitophagy. On the other hand, hypothermia increased the expression of Bcl-2 and Beclin-1 through disrupting the interaction between Bcl-2 and Beclin-1 during OGD/R, alleviating apoptosis and promoting mitophagy in the end. Thus, hypothermia provided neuroprotective effects via activating the PINK1/Parkin-VDAC3 signaling pathway.

Article Snippet: The materials used in this study were as follows: microglial mouse cell lines (catalog # CL-0493, Procell); high glucose Dulbecco’s modified Eagle’s medium (DMEM; catalog # C12430500BT, Gibco); 10% fetal bovine serum (FBS; catalog # 26010074, Gibco); DMEM without glucose (catalog # 11966025, Gibco); modular incubator chamber (catalog # MTC101, billups-rothenberg); single flowmeter (catalog # SFM3001, billups-rothenberg); dulbecco’s phosphate-buffered Saline (D-PBS, catalog # Lvn1023, livning); 0.4% trypan blue stain (catalog # C0040 solarbio); electron microscope fixing solution, (catalog #G1102, Servicebio); JC-1 MMP Assay (catalog # HY-K0601, medchemexpress); BBcellProbe M61 mPTP detection kit (catalog # BB-48122, BestBio); RIPA Lysis Buffer (catalog # C-1053, Applygen); Phenylmethanesulfonyl fluoride (PMSF; catalog # A1100, Applygen); Protease inhibitor cocktail (catalog # P1265-1, Applygen); NcmBlot blocking buffer (catalog # P30500, Ncmbio); rabbit anti-PINK1 antibody (catalog # 23274-1-AP, proteintech); rabbit anti-Parkin antibody (catalog # 32833, cell signaling technology); mouse anti-Parkin antibody (catalog # 4211, cell signaling technology); rabbit anti-VDAC3 antibody (catalog # 55260-1-AP, proteintech); rabbit anti-Bcl2 antibody (catalog # 26593-1-AP, proteintech); rabbit anti- Beclin-1 antibody (catalog # 3495 S, cell signaling technology); rabbit anti-Beta Actin antibody (catalog # 20536-1-AP, proteintech); rabbit anti-GAPDH antibody (catalog # 10494-1-AP, proteintech); rabbit anti-Cleaved Caspase 3 antibody (catalog # 9664, cell signaling technology); rabbit anti-caspase3 antibody (catalog # 14220, cell signaling technology); rabbit anti- SQSTM1/p62 antibody (catalog # 39749, cell signaling technology); rabbit anti-Cytochrom C antibody (catalog # 10993-1-AP, proteintech); rabbit anti-LC3B antibody (catalog # 43566, cell signaling technology); goat anti-rabbit HRP-conjugated IgG secondary antibody (catalog # ZB-2301, zsbio); Goat anti-Mouse IgG (H + L) Secondary Antibody, DyLightTM 488 (catalog # 35502, Thermo fisher scientific); Goat anti-Rabbit IgG (H + L) Secondary Antibody, DyLightTM 594(catalog # 35560, Thermo fisher scientific); HiScript III RT SuperMix for qPCR (+ gDNA wiper), (catalog # R323-01, Vazyme Biotech); FastPure Complex Cell/Tissue Total RNA Isolation Kit (catalog # RC113-C1, Vazyme Biotech); Taq Pro Universal SYBR qPCR Master Mix (catalog # Q712-03, Vazyme Biotech); QuickBlockTM Blocking Buffer for Immunol Staining (catalog # P0260, Beyotime).

Techniques: Expressing, Membrane

Journal: Scientific Reports

Article Title: Hypothermia regulates mitophagy and apoptosis via PINK1/Parkin-VDAC 3 signaling pathway during oxygen-glucose deprivation/recovery injury

doi: 10.1038/s41598-025-89176-w

Figure Lengend Snippet: Cellular signal pathway diagram of hypothermia at 4 h after temperature intervention. Under the condition of OGD/R, hypothermia inhibited Parkin recruitment to mitochondria and magnified mPTP opening via the high expression of VDAC3, and then aggravated the release of Cyt C through mPTP. Hypothermia also increased the expression of SQSTM/p62 and Beclin-1, but did not change the ratio of LC3B II/LC3B I. Finally, hypothermia induced apoptosis but did not promote mitophagy. The recruitment of Parkin from cytoplasm to mitochondria might be relevant to the overexpression of Bcl-2. (Cleaved PINK1, cPINK1; OMM, outer mitochondrial membrane; IMS, inner mitochondrial stroma; IMM, inner mitochondrial membrane). Red represents the different effects between 2 h and 4 h after hypothermia intervention.

Article Snippet: The materials used in this study were as follows: microglial mouse cell lines (catalog # CL-0493, Procell); high glucose Dulbecco’s modified Eagle’s medium (DMEM; catalog # C12430500BT, Gibco); 10% fetal bovine serum (FBS; catalog # 26010074, Gibco); DMEM without glucose (catalog # 11966025, Gibco); modular incubator chamber (catalog # MTC101, billups-rothenberg); single flowmeter (catalog # SFM3001, billups-rothenberg); dulbecco’s phosphate-buffered Saline (D-PBS, catalog # Lvn1023, livning); 0.4% trypan blue stain (catalog # C0040 solarbio); electron microscope fixing solution, (catalog #G1102, Servicebio); JC-1 MMP Assay (catalog # HY-K0601, medchemexpress); BBcellProbe M61 mPTP detection kit (catalog # BB-48122, BestBio); RIPA Lysis Buffer (catalog # C-1053, Applygen); Phenylmethanesulfonyl fluoride (PMSF; catalog # A1100, Applygen); Protease inhibitor cocktail (catalog # P1265-1, Applygen); NcmBlot blocking buffer (catalog # P30500, Ncmbio); rabbit anti-PINK1 antibody (catalog # 23274-1-AP, proteintech); rabbit anti-Parkin antibody (catalog # 32833, cell signaling technology); mouse anti-Parkin antibody (catalog # 4211, cell signaling technology); rabbit anti-VDAC3 antibody (catalog # 55260-1-AP, proteintech); rabbit anti-Bcl2 antibody (catalog # 26593-1-AP, proteintech); rabbit anti- Beclin-1 antibody (catalog # 3495 S, cell signaling technology); rabbit anti-Beta Actin antibody (catalog # 20536-1-AP, proteintech); rabbit anti-GAPDH antibody (catalog # 10494-1-AP, proteintech); rabbit anti-Cleaved Caspase 3 antibody (catalog # 9664, cell signaling technology); rabbit anti-caspase3 antibody (catalog # 14220, cell signaling technology); rabbit anti- SQSTM1/p62 antibody (catalog # 39749, cell signaling technology); rabbit anti-Cytochrom C antibody (catalog # 10993-1-AP, proteintech); rabbit anti-LC3B antibody (catalog # 43566, cell signaling technology); goat anti-rabbit HRP-conjugated IgG secondary antibody (catalog # ZB-2301, zsbio); Goat anti-Mouse IgG (H + L) Secondary Antibody, DyLightTM 488 (catalog # 35502, Thermo fisher scientific); Goat anti-Rabbit IgG (H + L) Secondary Antibody, DyLightTM 594(catalog # 35560, Thermo fisher scientific); HiScript III RT SuperMix for qPCR (+ gDNA wiper), (catalog # R323-01, Vazyme Biotech); FastPure Complex Cell/Tissue Total RNA Isolation Kit (catalog # RC113-C1, Vazyme Biotech); Taq Pro Universal SYBR qPCR Master Mix (catalog # Q712-03, Vazyme Biotech); QuickBlockTM Blocking Buffer for Immunol Staining (catalog # P0260, Beyotime).

Techniques: Expressing, Over Expression, Membrane

Journal: Cell reports

Article Title: Exploring the In Vivo Role of the Mitochondrial Calcium Uniporter in Brown Fat Bioenergetics

doi: 10.1016/j.celrep.2019.04.013

Figure Lengend Snippet:

Article Snippet: Rabbit polyclonal against VDAC , Cell Signaling Technologies , Cat#4866; RRID: AB_2272627.

Techniques: Synthesized, Recombinant, Saline, Protease Inhibitor, Lysis, Modification, Gene Expression, Enzyme-linked Immunosorbent Assay, Generated, Software

Journal: Molecular & Cellular Proteomics : MCP

Article Title: Interactome Analysis Identifies the Role of BZW2 in Promoting Endoplasmic Reticulum-Mitochondria Contact and Mitochondrial Metabolism

doi: 10.1016/j.mcpro.2023.100709

Figure Lengend Snippet:

Article Snippet: Antibody , Anti-VDAC2 (rabbit polyclonal) , Proteintech , 11663-1-AP , WB (1:1000).

Techniques: Cloning, Transfection, Transduction, Construct, Protease Inhibitor, Software, Calcium Assay, Bioassay, Isolation, Viability Assay, Sequencing, Modification, Western Blot, Bicinchoninic Acid Protein Assay