Fig. 4.

In vitro analysis of expression of pro-inflammatory mediators in primary cultures of differentiated human myotubes. (A) Purity of primary myotube cultures was assessed by immunohistochemical staining of the muscle cell marker desmin (Alexa-488; green) with a typical purity of 95% as revealed by comparison to phase contrast photos of the same culture well. Scale bar represents 100 µm. (B and C) mRNA-expression relative to GAPDH of chemokines (B) and cytokines (C) as assessed by quantitative (real-time) PCR in myotubes exposed 24 h to IFN-γ (300 U/ml), TNF-α (5 ng/ml) or IL-1β (10 ng/ml)—alone or in combinations. Data are given as mean + SD of two wells per condition from one representative experiment of four with similar results.