Fig. 5

The effect of cinacalcet on intracellular signaling for AMPK-eNOS, apoptosis, and oxidative stress in the podocytes cultured in low-glucose (LG; 5 mmol/l D-glucose) or high-glucose (HG; 30 mmol/l D-glucose) conditions with or without cinacalcet treatment (1 nM, 5 nM, 15 nM) (a–d). Representative Western blot analyses and quantitative analyses of total AMPK, phosphor-Thr¹⁷² AMPK, total eNOS, phospho-Ser¹¹⁷⁷ eNOS (a, *P < 0.05 and **P < 0.01 compared with LG control), SOD1 and SOD2 (b, *P < 0.05 and **P < 0.01 compared with LG control), dihydroethidium expression (c, *P < 0.05 and #P < 0.001 compared with other groups), Bcl-2, Bax, and TUNEL-positive podocytes (e, *P < 0.05 and **P < 0.01 compared with LG control and #P < 0.001 compared with other groups), and β-actin levels in the cultured podocytes and their quantitative analyses of the results are shown (n = 4 independent experiments in each experiments). d The effect of BAPTA-AM on cinacalcet-indueced in the podocytes cultured in low-glucose or high-glucose (HG; 30 mmol/l D-glucose) with or without cinacalcet treatment (15 nM). Representative Western blot analyses and quantitative analyses of CaMKKβ, phospho-LKB1, and total LKB1 (n = 4 independent experiments in each experiments). *P < 0.05 and **P < 0.01 compared with LG control. f The changes of intracellular signaling related to autophagy in podocytes exposed to cinacalcet and high-glucose media. Representative western blot analyses and quantitative analyses of beclin-1, LC3-II/LC3-I ratio, and β-actin levels in the cultured podocytes and their quantitative analyses of the results are shown (n = 4 independent experiments in each experiments). Representative immunofluorescent analyses of LC3 punctae in podocytes and the quantitative analyses of the results are shown. **P < 0.01 and # P < 0.001 compared with other groups (n = 6 independent experiments in each experiments)