Western blotting for iNOS and tubulin protein levels 24 h after treatment. Quantification represents the levels of iNOS protein normalized to tubulin and relative to the IFNγ/TNFα control.
RT–qPCR analysis of iNOS mRNA levels relative to the IFNγ/TNFα control 24 h after treatment.
Nitric oxide (NO) levels in the culture media of cells relative to the IFNγ/TNFα control 24 h after treatment.
C2C12 myoblast cells were transfected with control (siCTL) or AMPKα1 (siAMPKα‐1) targeting siRNA. Cells were subsequently treated with IFNγ/TNFα and the indicated doses of AICAR for 24 h. Western blotting for iNOS, tubulin, phospho‐Thr172 AMPKα (pAMPK), and AMPK protein levels. Quantification represents the levels of iNOS protein normalized to tubulin and relative to the siCTL, IFNγ/TNFα control.
Data information: All quantifications are of three independent experiments (
n = 3), and error bars represent the SEM. Significance between means was first determined using ANOVA. Significance
P‐values were calculated using Fisher's LSD. *
P < 0.05; **
P < 0.01 from equivalent siCTL samples;
††
P < 0.01 from IFNγ/TNFα‐treated controls.
Source data are available online for this figure.
