Journal: British Journal of Pharmacology

Article Title: Apocynin prevents cigarette smoking‐induced loss of skeletal muscle mass and function in mice by preserving proteostatic signalling

doi: 10.1111/bph.15482

Figure Lengend Snippet: Effect of CS exposure on tibialis anterior (TA) muscle weight, contractile performance and homeostatic changes. TA muscle weight (a), maximum contractile force (b), specific force at 120 Hz (c), and maximum contraction rate measurements (d) were analysed at the end of the experimental period. Quantitative PCR was performed to assess the expression of Igf1‐eb (e), Myostatin (f) and Tnfα (g) in homogenised TA muscle. Densitometry analysis of oxyblots for total oxidized proteins in the TA muscle (h). Data are expressed as mean ± SEM (n = 8–10 mice per group, Oxyblot analysis was conducted on n = 5 mice per group) from one of two independent experiments and analysed by two‐way ANOVA with multiple comparisons and Tukey post hoc test. * P < 0.05, significantly different from the relevant sham group; † P < 0.05, significantly different as indicated

Article Snippet: The suppliers of the materials used in these experiments are as follows: Winfield Red cigarettes (Phillip Morris, Australia); apocynin and DMSO (Sigma‐Aldrich, Australia); ketamine/xylazine and sodium pentobarbitone (Virbac Pty Ltd, Australia); acridine orange/ethidium bromide (Invitrogen, USA); Kwik‐Diff® reagent 1 fixative, High‐Capacity RNA‐to‐cDNA kit, pre‐developed TaqMan primers, cell culture reagents, Fluoromount‐GTM, with DAPI, murine IL‐6 ELISA Kit, PierceTM BCA Protein Assay Kit and SuperSignalTM West Femto Maximum Sensitivity Substrate for chemiluminescence detection (Thermo Fisher Scientific, USA); RNeasy Mini Kit (Qiagen, Germany); C2C12 murine myoblasts (American Type Culture Collection, USA; CRL‐1772); H 2 O 2 (Chem‐Supply, Australia); antibody for immunofluorescence (Santa Cruz Biotechnology, USA); MTS Cell Proliferation Assay (Promega, Australia); murine IGF‐1 DuoSet ELISA Kit (R&D Systems, USA); phosphorylation‐specific, actin antibodies and p62 for western blots (Cell Signaling Technology, USA); all other antibodies for western blots (Abcam, USA).

Techniques: Real-time Polymerase Chain Reaction, Expressing

Journal: British Journal of Pharmacology

Article Title: Apocynin prevents cigarette smoking‐induced loss of skeletal muscle mass and function in mice by preserving proteostatic signalling

doi: 10.1111/bph.15482

Figure Lengend Snippet: Effect of apocynin on C2C12 myotubes size and cellular stress. C2C12 myotubes were exposed to increasing concentrations of either H 2 O 2 or CSE with or without apocynin (500 nM) for 24 h. Changes in myotube diameters were quantified (a and f) from three independent experiments (n = 270 myotubes counted per condition). Quantitative PCR was performed to assess the expression of Cybb (b and g), Il‐6 (c and h), Igf1‐ea (d and i) and Igf1‐eb (e and j). Data are represented as mean ± SEM of three independent experiments (n = 7–9 per condition unless otherwise stated). * P < 0.05, significantly different from vehicle control (i.e., concentration 0); † P < 0.05, significantly different as indicated

Article Snippet: The suppliers of the materials used in these experiments are as follows: Winfield Red cigarettes (Phillip Morris, Australia); apocynin and DMSO (Sigma‐Aldrich, Australia); ketamine/xylazine and sodium pentobarbitone (Virbac Pty Ltd, Australia); acridine orange/ethidium bromide (Invitrogen, USA); Kwik‐Diff® reagent 1 fixative, High‐Capacity RNA‐to‐cDNA kit, pre‐developed TaqMan primers, cell culture reagents, Fluoromount‐GTM, with DAPI, murine IL‐6 ELISA Kit, PierceTM BCA Protein Assay Kit and SuperSignalTM West Femto Maximum Sensitivity Substrate for chemiluminescence detection (Thermo Fisher Scientific, USA); RNeasy Mini Kit (Qiagen, Germany); C2C12 murine myoblasts (American Type Culture Collection, USA; CRL‐1772); H 2 O 2 (Chem‐Supply, Australia); antibody for immunofluorescence (Santa Cruz Biotechnology, USA); MTS Cell Proliferation Assay (Promega, Australia); murine IGF‐1 DuoSet ELISA Kit (R&D Systems, USA); phosphorylation‐specific, actin antibodies and p62 for western blots (Cell Signaling Technology, USA); all other antibodies for western blots (Abcam, USA).

Techniques: Real-time Polymerase Chain Reaction, Expressing, Control, Concentration Assay