Fig. 7

Treatment with CamKII and proteasome inhibitors prevents NF-kB from being activated by high glucose. A rMC1 cells were pre-treated with 10 µM DMSO, Inh. XII, KT570 or BAPTA-AM for 2 h and with 500 nM epoxomicin for 1 h. Thereafter, compounds were washed out and 25 mmol/L glucose and low glucose/mannitol were added to culture media. After 20 min, whole cell lysates were prepared and analysed by Wb. Filters were probed with the anti-IkBα antibody. Total proteins stained by Ponceau S were used as loading control. Band intensity is reported in histograms. Data are presented as mean ± SD (n = 3); a representative blot is shown. Statistics was calculated by comparing each pair low–high glucose for any given compound assayed Multiple τ Student’s tests ***p < 0.001; The bands shown belong to the same gel, but this figure has been manipulated to clear out two lanes which were unnecessary for discussion (see uncropped files). B rMC1 cells were pre-treated with 10 µM DMSO and Inh. XII for 2 h and 500 nM epoxomicin for 1 h. Thereafter, compounds were washed out and 25 mmol/L glucose and low glucose or mannitol (controls) were added. After 40 min of stimulation nucleic acids were isolated and the expression of pro-inflammatory cytokines assayed by RT-PCR. β-actin was used as internal control. Histogram represents the relative quantity of transcripts calculated for any given cytokine for each experimental group (i.e., low glucose, high glucose and mannitol). Statistics was calculated for each cytokine separately. Data are presented as mean ± SD (n = 3). One-way ANOVA followed by Tukey’s post-hoc test (n = 3): *p < 0.05; ***p < 0.001