p ampk thr172 rabbit  (Cell Signaling Technology Inc)

Journal: Redox Biology

Article Title: Defective protein persulfidation is involved in obesity associated skeletal muscle dysfunction: role of SIRT-1

doi: 10.1016/j.redox.2025.103645

Figure Lengend Snippet: Effect of H 2 S donors on insulin resistance and inflammation in vitro in C2C12 myotubes exposed to the hyperlipidaemic environment. a-d Western blot analysis showing the expression and/or phosphorylation of AKT a,b and AMPK c,d in vehicle and SP-exposed C2C12 cells treated or not with NaHS or Erucin and stimulated with insulin. C2C12 unstimulated were used as basal control (CTL). Fold change data represent the mean ± SEM of four separate experiments. The blots shown are representative of four independent experiments with similar outcomes. The GAPDH bands confirm that similar amounts of proteins were loaded on the gel for each sample. e Bar graphs showing the mRNA expression levels of GLUT-4 in vehicle (n = 7) and SP-exposed C2C12 cells treated or not (n = 7) with NaHS (n = 4) or Erucin (n = 4) and then stimulated with insulin. f,g Bar graphs showing the mRNA expression levels of iNOS ( f ) and TNFα ( g ) in control myotubes (n = 4) and SP-exposed myotubes treated or not (n = 4) with NaHS (n = 4) or Erucin (n = 4) and then stimulated with insulin. h Bar graphs showing the mRNA expression levels of CBS, CSE and MPST in control myotubes and SP-exposed myotubes (n = 5) then stimulated with insulin. Data are expressed as 2 ∧−ΔΔct relative to β-actin for the results in e-h . All data are expressed as mean ± SEM and analyzed using one-way analysis of variance (ANOVA) followed by Bonferroni's post hoc test for the results in a-g and an unpaired-t-test two-tailed for the results in g . Differences are considered statistically significant when p was ≤0.05. Single asterisk (∗), double asterisk (∗∗) and triple asterisk (∗∗∗) denote a p-value of ≤0.05, 0.01, 0.001 vs vehicle, respectively. Single circle ( ○ ), double circle ( ○○ ) and triple circle ( ○○○ ) denote a p-value of ≤0.05, 0.01, 0.001 vs SP-exposed C2C12 group, respectively. Single ash ( # ) and triple ash ( ### ) denote a p-value ≤0.05, 0.01, 0.001 vs. unstimulated C2C12 (CTL).

Article Snippet: This was followed by overnight incubation at 4 °C with the primary antibodies: rabbit p -AKT antibody (Ser 473 1:1000; cat. no. 4060; Cell Signaling, Beverly, MA, USA); rabbit AKT antibody (1:12000; cat. no. 9272 Cell Signaling, Beverly, MA, USA); rabbit p -AMPK antibody (Thr 172 1:1000; cat. no. 50081; Cell Signaling, Beverly, MA, USA); mouse AMPK (1:1000; cat. no. 2793; Cell Signaling, Beverly, MA, USA); mouse SIRT-1 antibody (1:1000; cat. no. 60303-1-lg; Elabscience, USA).

Techniques: In Vitro, Western Blot, Expressing, Phospho-proteomics, Control, Two Tailed Test