voltage-dependent anion channel 1 (vdac1) antibody  (Santa Cruz Biotechnology)

Journal: International Journal of Oncology

Article Title: Targeting CALR reduces energy metabolism of esophageal cancer cells and inhibits tumor-associated fibroblast infiltration

doi: 10.3892/ijo.2025.5755

Figure Lengend Snippet: CALR regulates IP3R calcium ion release channels and ER stress. (A) Protein expression of (B) CALR, (C) CANX and (D) PDIA3 in KYSE150 after treatment with ER stress agonist thapsigargin. (E) Protein expression of (F) CALR, (G) CANX and (H) PDIA3 in KYSE410 cells following treatment with ER stress agonist thapsigargin. (I) In intracellular GRP78 expression after overexpression or knockdown of CALR in KYSE150. (J) Protein expression of GRP78 in KYSE150. (K) In intracellular GRP78 expression after overexpression or knockdown of CALR in KYSE410. (L) Protein expression of GRP78 in KYSE410. (M) Co-IP revealed the interaction of GRP75 and VDAC1 and IP3R1 in KYSE150. (N) Co-IP revealed the interaction of GRP75 and VDAC1 and IP3R1 in KYSE410. †† P<0.01, ††† P<0.001 vs. DMSO, ** P<0.01, *** P<0.001 vs. OE-NC, ### P<0.001 vs. sh-NC. CALR, Calreticulin; IP3R, inositol 1,4,5-Trisphosphate Receptor; ER, endoplasmic reticulum; CANX, calnexin; PDIA3, protein disulfide isomerase A3; ESCC, esophageal squamous cell carcinoma; GRP, glucose regulatory protein; VDAC, voltage-dependent anion channel; IB, immunoblotting; IP, immunoprecipitation; OE, overexpression; NC, negative control; sh, short hairpin.

Article Snippet: GRP75 (1:20,000; cat. no. 14887-1-AP), Voltage dependent anion channel 1 (VDAC1) (cat. no. 55259-1-AP, both Proteintech Group, Inc.) and inositol 1,4,5-Trisphosphate Receptor (IP3R1) (both 1:2,000; cat. no. DF3000, Affinity Biosciences).

Techniques: Expressing, Over Expression, Knockdown, Co-Immunoprecipitation Assay, Western Blot, Immunoprecipitation, Negative Control

Journal: Aging Cell

Article Title: The ultrastructural and proteomic analysis of mitochondria‐associated endoplasmic reticulum membrane in the midbrain of a Parkinson's disease mouse model

doi: 10.1111/acel.14436

Figure Lengend Snippet: Initial proteomic parsing of midbrain MAM fractions in controls and MPTP‐treated mice. (a) Subcellular fractions extracted from midbrain tissues were validated by specific organelle protein markers. Calnexin, Calreticulin, and Sigma‐1R were considered as consensus proteins for ER and MAM. VDAC1 and Cox IV were applied for mitochondria and MAM markers, and GAPDH was to prove cytosolic fractions. (b) Venn graph of identified proteins in each group ( n = 5 per group) and consensus MAM proteins. (c, d) Sample distribution plots by PCA (c) and PLS‐DA analysis (d) in MAM proteomics between controls and MPTP‐treated mice.

Article Snippet: Primary antibodies used in this study included that anti‐tyrosine hydroxylase (TH) rabbit mAb (1:2000, Cat. 58844, Cell Signaling Technology), anti‐glial fibrillary acidic portein (GFAP) rabbit mAb (1:2000, Cat. 80788, Cell Signaling Technology), anti‐glyceraldehyde‐3‐phosphate dehydrogenase (GAPDH) rabbit mAb (1:2000, Cat. 5174, Cell Signaling Technology), anti‐cytochrome‐c‐oxidase subunit 4 (Cox IV) rabbit Ab (1:2000, Cat. 5844, Cell Signaling Technology), anti‐sigma‐1 receptor (Sig‐1R) rabbit mAb (1:2000, Cat. 61994, Cell Signaling Technology), anti‐phosphatase and tensin homolog (PTEN) rabbit mAb (1:2000, Cat. 9188, Cell Signaling Technology), anti‐calreticulin rabbit mAb (1:2000, Cat. 92516, Abcam), anti‐calnexin (CNX) rabbit pAb (1:2000, Cat. ADI‐SPA‐860, Enzo Life Sciences), anti‐voltage dependent anion channel 1 (VDAC1) rabbit pAb (1:2000, Cat. 55259‐1‐AP, Proteintech), anti‐leucine‐rich repeat kinase 2 (LRRK2) rabbit mAb (1:1000, Cat. R380823, Zen‐bioscience), anti‐excitatory amino acid transporter 2 (EAAT2) rabbit mAb (1:1000, Cat. R381853, Zen‐bioscience), anti‐tropomyosin‐1 (TPM1) rabbit mAb (1:1000, Cat. R383145, Zen‐bioscience), and anti‐coatomer protein complex subunit zeta 1 (COPZ1) rabbit pAb (1:1000, Cat. 824631, Zen‐bioscience).

Techniques:

Journal: Journal of Thoracic Disease

Article Title: N-acetylcysteine, a small molecule scavenger of reactive oxygen species, alleviates cardiomyocyte damage by regulating OPA1 -mediated mitochondrial quality control and apoptosis in response to oxidative stress

doi: 10.21037/jtd-24-927

Figure Lengend Snippet: NAC regulates H9c2 cardiomyocyte apoptosis via OPA1 . H9c2 cardiomyocytes were treated with 250 µmol/L of H 2 O 2 for 12 h with or without 1 mmol/L of NAC pretreatment. (A-C) The release of cytochrome c from mitochondria to cytoplasm H9c2 cardiomyocytes was detected by western blot and normalized to VDAC and GAPDH, respectively; (D,E) H9c2 cardiomyocytes were double stained with Annexin V-FITC and PI-PE, and the apoptosis rate was measured using flow cytometry. The mitochondrial apoptosis pathway was verified by measuring (F) H9c2 cardiomyocyte caspase-3 activity levels. Statistical comparisons were performed using a one-way ANOVA. *, P<0.05 vs . control group; # , P<0.05 vs . H 2 O 2 group; & , P<0.05 vs . NAC group. VDAC, voltage-dependent anion channel; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; NAC, N-acetylcysteine; V-FITC, V-fluorescein isothiocyanate; PI, propidium iodide; PE, phycoerythrin; ANOVA, analysis of variance.

Article Snippet: The following primary antibodies were used to incubate the membranes: anti-Cytochrome C (Cat No: 66264-1-Ig; dilution: 1:1,000, Proteintech, Rosemont, IL, USA), anti-voltage-dependent anion channel (VDAC) (Cat No: 66345-1-Ig; dilution: 1:1,000, Proteintech), or anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (Cat No: 60004-1-Ig; dilution: 1:10,000, Proteintech) at 4 °C overnight.

Techniques: Western Blot, Staining, Flow Cytometry, Activity Assay, Control