Figure 9.

Effect of PIF on cytoplasmic I-κBα (A) and nuclear-bound NF-κB (C) determined 30 min after PIF addition to murine myotubes transfected with constitutively active (pCO₂ □) and mutant (pKS1 ▪) PKC_α. (A) Myotubes transfected with either pCO₂ (lanes 1–6) or pKS1 (lanes 7–12) were treated with 0 (lanes 1 and 7), 1.05 (lanes 2 and 8), 2.1 (lanes 3 and 9), 4.2 (lanes 4 and 10), 10 (lanes 5 and 11) or 16.8 nM PIF (lanes 6 and 12). The densitometric analysis is an average of three replicate blots. Differences from 0 nM PIF are shown as ^cP<0.001. (B) Actin loading control for the blot shown in (A). (C) EMSA of NF-κB nuclear binding in murine myotubes transfected with pCO₂ (lanes 2 and 6) and pK1 (lanes 7–11). Myotubes were treated with 0 (lanes 2 and 7), 2.1 (lanes 3 and 8), 4.2 (lanes 4 and 9), 10 (lanes 5 and 10) and 16.8 nM PIF (lanes 6 and 11). Lane 1 is a negative control containing the labelled probe without a nuclear extract; lane 12 contains a 100-fold excess of unlabelled NF-κB probe and lane 13 is a positive control for NF-κB (supplied by the manufacturers of the kit). The densitometric analysis is an average of three replicate EMSAs. Differences from control are indicated as ^bP<0.01 and ^cP<0.001.