Fig. 3

A second cycle of NF-kB activation by high glucose occurs through a typical signalling pathway. Whole cell lysates were prepared from rMC1 cells challenged with 25 mmol/L glucose and with low glucose or mannitol for 1, 2 and 3 h and analysed by Wb. Filters were probed with antibodies anti-pIkBα(Ser32), -IkBα, -pIKKβ(Ser80), -IKKβ, -p65(Ser536), and -p65. The pIkBα(Ser32)/IkBα and p65(Ser536)/p65 ratios are shown. Being pIKKβ undetectable in control cells, the unphosphorylated/phosphorylated ratio of this protein is not reported. Total proteins stained by Ponceau S were used as loading control. Histograms represent the results of densitometric analysis of the immunoreactive bands; data are the mean ± SD of three independent experiments; a representative blot is shown. Statistics were calculated for high glucose vs low glucose treated cells using a One-way ANOVA followed by Tukey’s post-hoc test: *p < 0.05; ***p < 0.001