Fig. 2

Inflammatory priming provokes the expression of high levels of several chemokines and iNOS by C2C12 cells. (A-F) Expression levels of chemokines and iNOS in C2C12 cells stimulated with murine IFN-γ and TNF-α (10 ng/ml each) for 24 h were assayed by qRT-PCR (n = 3). (G) The protein expression of iNOS and β-actin (loading control) were determined by immunoblotting in C2C12 cells stimulated with murine IFN-γ and TNF-α (10 ng/ml each) for 24 h (n = 3). The original images of blots are in the Supplementary Information. (H) C2C12 cells were stimulated with murine IFN-γ and TNF-α (10 ng/ml each) for 24 h and the supernatants were assayed for nitrate by a modified Griess reagent (n = 3). Values are presented as mean ± SEM. Statistical analysis was performed by two-tailed unpaired t test.