Figure 4
EDN1 and EDNRB protein expression and cell viability in 661W cultured cells under glucose-free conditions. (a–d) The relative amount of live cells was measured using WST-8 following 48 h treatment with DMEM/high glucose media or DMEM/glucose-free media with or without KUS121 (100 µM). (a) Quantitative analysis of live cells (n = 3, for all treatments). 661W cells were cultured with high glucose [labeled ‘‘(-)’’] or treated in glucose-free media with (labeled ‘‘K121’’) or without (control, labeled ‘‘C’’) KUS121. (b–d) Representative images of 661W cells cultured under each condition. The black bar represents 50 µm. (e–h) Protein expression of EDN1 (e,g) and EDNRB (f,h) in 661W cells was analyzed by western blotting. 661W cells were cultured with high glucose media [labeled ‘‘(-)’’] or with glucose-free media with (labeled ‘‘K’’) or without (control, labeled ‘‘C’’) KUS121 for 24 h before the analysis. Actin was used as a loading control. Complete scans of western blots are shown in Supplementary Fig. S2b,c. Relative expression of EDN1 (g) and EDNRB (h) was shown as a ratio to actin (n = 4, for both treatments). *p < 0.05, **p < 0.01 and ***p < 0.001, compared with the control (in a and g) or to the cells cultured in high glucose media (in h), Tukey’s HSD.
