Journal: Biology

Article Title: N-acetyl-L-cysteine Prevents Lactate-Mediated PGC1-alpha Expression in C2C12 Myotubes

doi: 10.3390/biology8020044

Figure Lengend Snippet: qPCR analysis of PGC1-alpha and SIRT1 expression in C2C12 myotubes cultured with L-lactate. C2C12 myotubes were cultured for 2 ( a ) and 6 h ( b ) with L-lactate at different concentrations (0, 10, 30, and 50 mM). In the time between lactate culture and sample collection (one hour), cells were cultured with DM. Error bars show standard deviations. * p < 0.05 from 0 mM. All the experiments were performed three independent times with three replicates each time.

Article Snippet: Membranes were then probed at 4 °C overnight in TBS-T containing 0.4% NaN3 and 1:1000 dilutions of specific antibodies against PGC1-alpha (Novus Biologicals, Centennial, CO, USA) and GAPDH (Abcam, Cambridge, UK).

Techniques: Expressing, Cell Culture

Journal: Biology

Article Title: N-acetyl-L-cysteine Prevents Lactate-Mediated PGC1-alpha Expression in C2C12 Myotubes

doi: 10.3390/biology8020044

Figure Lengend Snippet: qPCR analysis of PGC1-alpha, Cyt C, and SIRT1 in C2C12 myotubes cultured with lactate and N-acetyl-L-cysteine (Nac). C2C12 myotubes were cultured for 2 h with L-lactate (0 and 50 mM) in the presence or absence of Nac (2 mM). Samples for gene expression analysis were obtained at 0 ( a ) and 3 h ( b ) after the culture with lactate and Nac. In the time between lactate culture and sample collection (0 to 3 h), cells were cultured with DM. Error bars show standard deviations. * p < 0.05 from 0 mM. All the experiments were performed three independent times with three replicates each time.

Article Snippet: Membranes were then probed at 4 °C overnight in TBS-T containing 0.4% NaN3 and 1:1000 dilutions of specific antibodies against PGC1-alpha (Novus Biologicals, Centennial, CO, USA) and GAPDH (Abcam, Cambridge, UK).

Techniques: Cell Culture, Gene Expression

Journal: Biology

Article Title: N-acetyl-L-cysteine Prevents Lactate-Mediated PGC1-alpha Expression in C2C12 Myotubes

doi: 10.3390/biology8020044

Figure Lengend Snippet: qPCR analysis for intermittent and continuous exposure to lactate in C2C12 myotubes and Western blot of PGC1-alpha after continuous or intermittent exposure to lactate. C2C12 myotubes were cultured with different concentration patterns of L-lactate (see ). Samples for qPCR analysis were obtained 0 ( a ) and 3 h ( b ) after the exposure series. Samples for Western blot ( c ) were obtained 3 h after the exposure series (see ). In the time between lactate culture and sample collection (zero or three hours), cells were cultured with DM. Error bars show standard deviations. * p < 0.05 from non-lactate exposure (0 × 0). RNA experiments were performed three independent times with three replicates each time, and protein experiments with three independent samples.

Article Snippet: Membranes were then probed at 4 °C overnight in TBS-T containing 0.4% NaN3 and 1:1000 dilutions of specific antibodies against PGC1-alpha (Novus Biologicals, Centennial, CO, USA) and GAPDH (Abcam, Cambridge, UK).

Techniques: Western Blot, Cell Culture, Concentration Assay