Fig. 6

G6PD siRNA abrogates the glucose potentiation of IL1β-evoked pro-inflammatory response. (a) G6PD levels, determined by Western blot, in cells treated with scrambled siRNA or G6PD siRNA and submitted for 18 h IL1β (10 ng/mL) in medium containing 5.5 or 22 mmol/L glucose. (b) NADPH oxidase activity, determined by lucigenin-derived chemiluminescence in cells treated with scrambled siRNA and G6PD siRNA and exposed to IL1β (10 ng/mL) during 18 h of incubation, in medium initially containing 5.5 or 22 mmol/L glucose. Results are the mean ± standard error of 3–6 separate experiments expressed as percentage of the relative light units produced by 10 ng/mL IL1β in control cells incubated in a medium initially containing 5.5 mmol/L glucose (380.1 ± 59.7 RLU/µg protein min⁻¹). (c and d) iNOS levels, determined by Western blot, in cells untreated or treated with scrambled (scr)-siRNA and G6PD siRNA and exposed to IL1β (10 ng/mL) during 18 h of incubation in medium initially containing 5.5 or 22 mmol/L glucose. The gels and blots are representative of 3–5 separate experiments, while the bars are expressed as percentage of the activation or the expression produced by treatment with IL1β in sc-siRNA cells incubated in a medium with 5.5 mmol/L glucose. *P < 0.05 vs respective control (c). ^† P < 0.05 vs respective value in 5.5 mmol/L glucose. ^# P < 0.05 vs respective value in scr-siRNA