Sirolimus Enhances Cyclosporine A-Induced Cytotoxicity in Human Renal Glomerular Mesangial Cells
Figure 3
CsA/SRL cotreatment caused a biphasic increase in ERK 1/2 activity in HMCs. HMCs were grown to confluency and treated with vehicle, 4.2 μM CsA, 1 μM SRL, or CsA/SRL 4.2 μM CsA + 1 μM SRL. Phosphorylated ERK 1/2 MAPK or whole-cell ERK 1/2 MAPK was probed by Western blotting. Representative blots from one of three independent experiments are shown. (a) ERK 1/2 phosphorylation following CsA treatment, (b) ERK 1/2 phosphorylation following SRL treatment, (c) ERK 1/2 phosphorylation following CsA/SRL cotreatment. (d) ERK 1/2 phosphorylation in the presence of vehicle (fresh medium was applied to cells at t0). Densitometric analysis was conducted by calculating the ratio of phosphorylated ERK 1/2 compared to total whole-cell ERK 1/2 and then comparing this ratio to time-matched vehicle controls. Each column represents the mean ± SEM of a minimum of 3 independent experiments performed in duplicate. Data was analysed by ANOVA and comparisons between control and multiple treatment groups were made using the Dunnet posttest. *Indicates statistical difference compared to control. *, **.
