mitochondrial protein extraction kit  (Sangon Biotech)

Journal: Human Mutation

Article Title: NDUFA4 Deletion Upregulates VDAC1 to Promote Mitochondrial Damage, Endoplasmic Reticulum Expansion, and Neuronal Apoptosis

doi: 10.1155/humu/8889386

Figure Lengend Snippet: Effects of NDUFA4 knockout on VDAC1 expression and mitochondrial‐related cellular changes. Cerebellum tissues from four pairs of wild‐type and NDUFA4 knockout mice were collected. (A) NDUFA4 and VDAC1 expression detected by Western blot. (B) Activity of ETC Complex IV measured using the MitoCheck Complex IV Activity Assay Kit. (C) Western blot analysis of cytochrome C expression in mitochondrial and cytoplasmic fractions. (D) Western blot analysis of apoptosis‐related proteins (caspase‐3, cleaved caspase‐3, caspase‐12, Bax, and Bcl‐2) and ER stress–related proteins (CHOP, ATF4, PERK, and Eif2 α ). ( n = 4). ∗ p < 0.05, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001.

Article Snippet: Cytoplasmic and mitochondrial proteins were isolated using the Cytoplasmic and Mitochondrial Protein Extraction Kit (C500051‐0050, Sangon Biotech).

Techniques: Knock-Out, Expressing, Western Blot, Activity Assay

Journal: Bioactive Materials

Article Title: Mechanism and regulatory strategy study on promoting vascularized bone regeneration via intracellular zinc ion transport

doi: 10.1016/j.bioactmat.2025.07.020

Figure Lengend Snippet: BMSC mitochondrial permeability increased and cytoplasmic Zn 2+ accumulated in the hypoxic microenvironment (a) TEM results of BMSCs cultured in a hypoxic milieu for different time periods. (b) Western blotting detecting the expression of MFN1 in the mitochondrial protein (up) and the expression of Cyt-C in the cytoplasmic protein (bottom). (c) mPTP detection results. (d) Multiple metal ion detection in the cytoplasmic components of BMSCs during the process of hypoxia induction. (e) Flow cytometry detection of cytoplasmic Zn 2+ in BMSCs under hypoxia induction. (f,g) Fluorescence colocalization of cellular mitochondria, endoplasmic reticulum, and cytoplasmic Zn 2+ . (h,i) Flow cytometry detection of exogenous or endogenous Zn 2+ in BMSCs. (j) The BCA quantitative results of the secreted protein in CM. (k) ELISA results of MMP-10 expression in BMSC CM. All in vitro results are representative of at least three independent experiments (n = 3 for each experiment). The yellow rectangle in (a) indicates the site of endoplasmic reticulum-mitochondria coupling. Scale bar in (f) & (g) = 10 μm. Data are presented as mean ± SD. ∗p < 0.05. ∗∗p < 0.01. Ctrl = Control. CM = conditioned medium. BMSC = bone marrow mesenchymal stem cell.

Article Snippet: Mitochondrial proteins were extracted using a mitochondrial protein extraction kit (Solarbio, Beijing, China), according to the manufacturer's instructions.

Techniques: Permeability, Cell Culture, Western Blot, Expressing, Flow Cytometry, Fluorescence, Enzyme-linked Immunosorbent Assay, In Vitro, Control

Journal: Journal of Functional Foods

Article Title: Pulsed electric field-treated malvacin-3-O-galactoside and whey protein complexes alleviate silica-induced pulmonary fibrosis by promoting cellular mitochondrial autophagy and inhibiting cellular pyroptosis

doi: 10.1016/j.jff.2025.106895

Figure Lengend Snippet: Fig. 5. PEF-M3G-WP promotes cellular autophagy to attenuate apoptosis in silicosis mouse lungs. (A–D) WB analysis of the relative expression of PINK1 and Parkin proteins in mitochondrial lysates of mouse lung tissues (n = 3). (E–J) WB analysis of the relative expression of NLRP3, CASP1 p20 and GSDMD NT proteins in lung tissues (n = 4). (K–L) q-PCR analysis of mRNA content of autophagy genes PINK1 and Parkin in lung tissues (n = 3–5). (M–O) q-PCR analysis of mRNA content of lung tissue pyroptosis genes Nlrp3, Casp1 p20, and Gsdmd (n = 4). All data are expressed as mean ± SEM (n = 3). Different lowercase letters indicate significant dif ferences (P < 0.05).

Article Snippet: Mitochondrial proteins were isolated using a Mitochondrial Protein Extraction Kit (WLA034, Wanleibio, China).

Techniques: Expressing

Journal: Journal of Functional Foods

Article Title: Pulsed electric field-treated malvacin-3-O-galactoside and whey protein complexes alleviate silica-induced pulmonary fibrosis by promoting cellular mitochondrial autophagy and inhibiting cellular pyroptosis

doi: 10.1016/j.jff.2025.106895

Figure Lengend Snippet: Fig. 7. PEF-M3G-WP activates mitochondrial autophagy genes to inhibit AM pyroptosis. (A–B) q-PCR analysis of the relative mRNA content of autophagy genes P62 and Atg5 in MH-S cell lysates (n = 4). (C–D) q-PCR analysis of the relative mRNA content of pyroptosis genes IL18 and IL1b in MH-S cell lysates (n = 5). All data are expressed as mean ± SEM (n = 3). Different lowercase letters indicate significant differences (P < 0.05).

Article Snippet: Mitochondrial proteins were isolated using a Mitochondrial Protein Extraction Kit (WLA034, Wanleibio, China).

Techniques:

Journal: FASEB BioAdvances

Article Title: Protective Effects of Rat Bone Marrow Mesenchymal Stem Cells‐Derived Fusogenic Plasma Membrane Vesicles Containing VSVG Protein Mediated Mitochondrial Transfer on Myocardial Injury In Vitro

doi: 10.1096/fba.2024-00235

Figure Lengend Snippet: Expression of mitochondrial function‐related genes/proteins and fPMVs‐related proteins determined by quantitative real‐time PCR (qPCR) or western blot. (A) The expression of MT‐ND4 , ACTB , and HDAC1 in the rBMSCs and rBMSCs with VSVG overexpression. * p < 0.05, vs. rBMSCs. (B) The expression of MT‐ND4 , ACTB , and HDAC1 in the fPMVs and fPMVs with VSVG overexpression. * p < 0.05, vs. fPMVs. (C) The protein expression of VSVG, CD81, eEF2, MTCO1, and TOMM20 in the rBMSCs and rBMSCs with VSVG overexpression. * p < 0.05, vs. rBMSCs. (D) The protein expression of VSVG, CD81, eEF2, MTCO1, and TOMM20 in the fPMVs and fPMVs with VSVG overexpression. * p < 0.05, vs. fPMVs.

Article Snippet: The H9c2 cells with different treatments were employed to isolate mitochondria using a Mitochondrial Isolation and Protein Extraction Kit (Proteintech, Wuhan, China) according to the recommendations of the manufacturer.

Techniques: Expressing, Real-time Polymerase Chain Reaction, Western Blot, Over Expression

Journal: FASEB BioAdvances

Article Title: Protective Effects of Rat Bone Marrow Mesenchymal Stem Cells‐Derived Fusogenic Plasma Membrane Vesicles Containing VSVG Protein Mediated Mitochondrial Transfer on Myocardial Injury In Vitro

doi: 10.1096/fba.2024-00235

Figure Lengend Snippet: Effects of fPMVs on the mitochondrial functions of H9c2 cells induced by HR. (A) The expression of MT‐ND1 in the HR‐induced H9c2 cells treated with fPMVs. * p < 0.05, vs. control. The contents of TNF‐α (B), IL‐1β (C), ATP (D), and MRCC IV (E) in the cellular mitochondria with different treatments. (F) The ROS levels in the cellular mitochondria with different treatments under a laser scanning confocal microscope. * p < 0.05, vs. control, # p < 0.05, vs. HR.

Article Snippet: The H9c2 cells with different treatments were employed to isolate mitochondria using a Mitochondrial Isolation and Protein Extraction Kit (Proteintech, Wuhan, China) according to the recommendations of the manufacturer.

Techniques: Expressing, Control, Microscopy