Journal: Physiological reports

Article Title: High-fat diet increases electron transfer flavoprotein synthesis and lipid respiration in skeletal muscle during exercise training in female mice.

doi: 10.14814/phy2.15840

Figure Lengend Snippet: FIGURE 8 Twelve weeks of high-fat diet induced mitochondrial remodeling through fission, fusion, and BNIP3-dependent autophagy and mitophagy mechanisms. (a) markers of mitochondrial dynamics (fission and fusion) and (b) representative blots. (c) whole-cell autophagy and (d) representative blots. (e) markers of mitophagy in mitochondrial subfractions (subsarcolemmal [SSM] and intermyofibrillar [IMFM]) and (f) representative blots. Whole-cell lysates are from quadriceps and isolated mitochondria are from gastrocnemius. Data displayed as mean ± SD. Effects of diet and exercise were evaluated using two-way ANOVA. Images on representative blots come from separate membranes and channels.

Article Snippet: Secondary antibodies were diluted in 5% BSA-TBST or 5% nonfat dry milk-TBST and membranes incubated at room temperature for 1 h. Primary antibodies were diluted 1:1000 and purchased from Abcam (Cambridge, United Kingdom) for OXPHOS cocktail (110413), ETFα (110316), ETFβ (240593), Trimethylated ETFβ (76118), ETFDH (103910), CPT1 (134988), PINK1 (23707), Mitochondrial fission factor (MFF) (81127); Cell signaling (Danvers, MA) for LC3 (12741), p62 (39749), Parkin (4211), and BNIP3 (3769), OPA1 (80471), mitofusion 2 (MFN2) (9482), Abcam, DRP1 (14647); Other primary antibodies included METTL20 (87995, Novus Biologicals, Littleton, CO), HADH (PA5-28203, Thermo Fisher Scientific, Waltham, MA), and BCL2 (7382, Santa Cruz Biotechnology, Dalla, TX).

Techniques: Isolation

Journal: Journal of Cellular and Molecular Medicine

Article Title: Platinum‐based combination chemotherapy triggers cancer cell death through induction of BNIP3 and ROS, but not autophagy

doi: 10.1111/jcmm.14898

Figure Lengend Snippet: Increased levels and mitochondria localization of BNIP3 were identified upon treatment with anticancer drugs. A, B, A549 cells (2 × 10 5 ) were treated with cisplatin (2 μg/mL), LBH589 (100 nmol/L), or rapamycin (100 nmol/L), or a combination of two drugs for 24 h (A) or 48 h (B). The mRNA or protein levels of BNIP3 were detected by qRT‐PCR (A) or Western blot (B), respectively. C, A549 cells were transfected with pEGFP‐BNIP3 plasmid. Sixteen hours later, cells were treated with anticancer drugs for 24 h. Fluorescent dye‐labelled mitochondria were detected using fluorescence microscopy upon treatment of 2 nmol/L of tetramethylrhodamine methyl ester perchlorate (TMRM) for 30 min. Results are representative of at least three independent experiments. ** P < .01 compared with untreated cells

Article Snippet: Rapamycin, LC3 antibody and BNIP3 antibody were obtained from Calbiochem, Novus Biologicals and Abcam, respectively.

Techniques: Quantitative RT-PCR, Western Blot, Transfection, Plasmid Preparation, Fluorescence, Microscopy

Journal: Journal of Cellular and Molecular Medicine

Article Title: Platinum‐based combination chemotherapy triggers cancer cell death through induction of BNIP3 and ROS, but not autophagy

doi: 10.1111/jcmm.14898

Figure Lengend Snippet: Anticancer drugs induced mitochondrial membrane insertion of BNIP3, resulting in the generation of ROS and cell death. A, A549 cells (2 × 10 5 ) were transfected with pcDNA empty vector or ΔTM‐BNIP3 plasmid. Sixteen hours later, cells were treated with cisplatin (2 μg/mL), LBH589 (100 nmol/L), or rapamycin (100 nmol/L), or a combination of two drugs for 48 h. Cell viability was analysed using a Celltiter‐Glo luminescent cell viability assay. B, A549 cells were treated with cisplatin (2 μg/mL), LBH589 (100 nmol/L), or rapamycin (100 nmol/L), or a combination of two drugs for 48 h. Cells were treated with or without the ROS inhibitor NAC (5 mmol/L) 24 h prior to analysis. Cell viability was analysed using a Celltiter‐Glo luminescent cell viability assay. Results are representative of at least three independent experiments. * P < .05 and ** P < .01

Article Snippet: Rapamycin, LC3 antibody and BNIP3 antibody were obtained from Calbiochem, Novus Biologicals and Abcam, respectively.

Techniques: Membrane, Transfection, Plasmid Preparation, Cell Viability Assay

Journal: Journal of Cellular and Molecular Medicine

Article Title: Platinum‐based combination chemotherapy triggers cancer cell death through induction of BNIP3 and ROS, but not autophagy

doi: 10.1111/jcmm.14898

Figure Lengend Snippet: Inhibition of autophagy augments chemotherapeutic drug‐induced cell death. A, B, A549 cells (2 × 10 5 ) were treated with cisplatin (2 μg/mL), LBH589 (100 nmol/L), or rapamycin (100 nmol/L), or a combination of two drugs for 48 h. Cells were treated with or without 3‐MA (3 mmol/L) 1 h prior to analysis. A, Cell viability was analysed with a Celltiter‐Glo luminescent cell viability assay. B, Cells were stained with propidium iodide (10 μg/mL) for 30 min. The cell cycle was determined using flow cytometry. C, A549 cells were transfected with pcDNA empty vector or BNIP3 plasmid. Sixteen hours later, A549 cells were treated with chemotherapeutic drugs with or without 3‐MA (3 mmol/L) 1 h prior to analysis. Cell viability was analysed with a Celltiter‐Glo luminescent cell viability assay. Results are representative of at least three independent experiments. * P < .05 and ** P < .01

Article Snippet: Rapamycin, LC3 antibody and BNIP3 antibody were obtained from Calbiochem, Novus Biologicals and Abcam, respectively.

Techniques: Inhibition, Cell Viability Assay, Staining, Flow Cytometry, Transfection, Plasmid Preparation

Journal: Journal of Cellular and Molecular Medicine

Article Title: Platinum‐based combination chemotherapy triggers cancer cell death through induction of BNIP3 and ROS, but not autophagy

doi: 10.1111/jcmm.14898

Figure Lengend Snippet: A proposed model for the role of BNIP3 and autophagy in chemotherapeutic drug‐induced cell death. Anticancer drugs augment Binp3 and autophagy in lung cancer cells. Anticancer drugs induction of mitochondrial membrane insertion of BNIP3 may promote ROS and potential membrane loss, resulting in apoptotic or necrotic cell death. However, anticancer drug‐induced autophagy may also be involved in cell survival

Article Snippet: Rapamycin, LC3 antibody and BNIP3 antibody were obtained from Calbiochem, Novus Biologicals and Abcam, respectively.

Techniques: Membrane