Figure 5
AGE-BSA increases cell proliferation by activating KCa3.1 channels in rat VSMCs. (a) Mean values of cell counting in cells treated without (control) or with BSA (200 μg/ml), AGE-BSA (200 μg/ml), and AGE-BSA plus 100 nM TRAM-34 or 5 μg/ml anti-RAGE antibody 24 h (n=6, **P<0.01 vs control or BSA, ##P<0.01 vs AGE-BSA alone). (b) Immunofluorescent BrdU-staining images ( × 100) of the nuclei in VSMCs treated with the interventions as described in a. Nuclei counterstained in blue (4,6-diamidino-2-phenylindole) and BrdU-positive nuclei shown in pink. No visible pink color (the color of rhodamine-labeled second antibody) was observed in negative control without incubation of the primary antibody. (c) Mean percent values of BrdU-incorporated nuclei in VSMCs treated with the interventions as described in a. (n=6, **P<0.01 vs control or BSA; ##P<0.01 vs AGE-BSA alone). (d) Mean values of cell number counted in cells transfected with control siRNA or KCa3.1 siRNA (100 nM) and treated with BSA or AGE-BSA (200 μg/ml) for 24 h (n=6, **P<0.01 vs control siRNA with BSA; ##P<0.01 vs control siRNA with AGE-BSA). (e) BrdU-incorporation staining ( × 100) in cells treated with the interventions as described in d. (f) Mean percentage of values of proliferation cells with pink color in cells treated with the interventions as described in d (n=6, **P<0.01 vs control siRNA with BSA; ##P<0.01 vs control siRNA with AGE-BSA).
