Volume 54, Issue 2, Pages 492-508 (August 1998) Corticosterone and 11-dehydrocorticosterone stimulate Na,K-ATPase gene expression in vascular smooth muscle cells  Shigeaki Muto, Jun Nemoto, Satoru Ebata, Kiyoshi Kawakami, Yasushi Asano  Kidney International  Volume 54, Issue 2, Pages 492-508 (August 1998) DOI: 10.1046/j.1523-1755.1998.00033.x Copyright © 1998 International Society of Nephrology Terms and Conditions

Figure 1 Effects of 12-hour incubation with corticosterone (CS), 11-dehydrocorticosterone (DHCS), or 21-desoxycortisone (DC) on Na,K-ATPase α1- and β1-mRNA accumulation in vascular smooth muscle cells (VSMC). (A) Northern blot analyses. (B) Quantitations of the Northern blot. Data are means ± se of 3 separate experiments, expressed as a relative increase in mRNA expression compared with control samples.*P < 0.005, **P < 0.001 compared to control. Kidney International 1998 54, 492-508DOI: (10.1046/j.1523-1755.1998.00033.x) Copyright © 1998 International Society of Nephrology Terms and Conditions

Figure 2 Time course of Na,K-ATPase α1- and β1-mRNA accumulation by CS in VSMC. (A) Northern blot analyses. (B) Quantitations of the Northern blot. Abundance of mRNA is expressed as units of densitometry relative to time 0, and each data point is means ± se of five separate experiments.*P < 0.05, #P < 0.01, †P < 0.001 compared to control. Symbols are: (▪) control; (♦) CS. Kidney International 1998 54, 492-508DOI: (10.1046/j.1523-1755.1998.00033.x) Copyright © 1998 International Society of Nephrology Terms and Conditions

Figure 3 Time course of Na,K-ATPase α1- and β1-mRNA accumulation by DHCS in VSMC. (A) Northern blot analyses. (B) Quantitations of the Northern blot. Abundance of mRNA is expressed as units of densitometry relative to time 0, each point is means ± se of five separate experiments.*P < 0.01, #P < 0.005, †P < 0.001 compared to control. Symbols are: (▪) control; (♦) DHCS. Kidney International 1998 54, 492-508DOI: (10.1046/j.1523-1755.1998.00033.x) Copyright © 1998 International Society of Nephrology Terms and Conditions

Figure 4 Dose-dependent effects of CS and DHCS on Na,K-ATPase α1- and β1-mRNA accumulation in VSMC.(A) Northern blot analyses for CS- or DHCS-supplemented media. (B) Quantitations of the Northern blot. Abundance of mRNA is expressed as units of densitometry relative to control, and data are means ± se of 4 separate experiments.*P < 0.05, #P < 0.01,†P < 0.001 compared to control. Kidney International 1998 54, 492-508DOI: (10.1046/j.1523-1755.1998.00033.x) Copyright © 1998 International Society of Nephrology Terms and Conditions

Figure 5 Effects of a transcription inhibitor, actinomycin D (ACD), on CS- and DHCS-induced upregulation of α1- and β1-mRNA accumulation in VSMC. (A) Northern blot analyses in VSMC exposed to control MEM, CS- or DHCS-supplemented MEM in the absence or presence of 5 μg/ml ACD. (B) Quantitations of the Northern blot for CS- or DHCS-supplemented media. Data are means ± se of 4 separate experiments, expressed as a relative increase in mRNA expression compared with control samples. Kidney International 1998 54, 492-508DOI: (10.1046/j.1523-1755.1998.00033.x) Copyright © 1998 International Society of Nephrology Terms and Conditions

Figure 6 Effects of the GR antagonist, RU38486 (RU), on CS- and DHCS-induced up-regulation of α1- and β1-mRNA in VSMC.(A) Northern blot analyses in VSMC exposed to control MEM, CS- or DHCS-supplemented MEM in the absence or presence of 10-5 m RU. (B) Quantitations of the Northern blot. Data are expressed as a relative increase in mRNA expression compared with control samples. The number of experiments done on CS- or DHCS-treated VSMC is 4. Kidney International 1998 54, 492-508DOI: (10.1046/j.1523-1755.1998.00033.x) Copyright © 1998 International Society of Nephrology Terms and Conditions

Figure 7 Effects of the MR antagonist, ZK91587 (ZK), on CS- and DHCS-induced up-regulation of α1- and β1-mRNA in VSMC. (A) Northern blot analyses in VSMC exposed to control MEM, CS- or DHCS-supplemented MEM in the absence or presence of 10-5 m ZK. (B) Quantitations of the Northern blot. Data are expressed as a relative increase in mRNA expression compared with control samples. The number of experiments done on CS- or DHCS-treated VSMC is 5. Kidney International 1998 54, 492-508DOI: (10.1046/j.1523-1755.1998.00033.x) Copyright © 1998 International Society of Nephrology Terms and Conditions

Figure 8 Effects of the 11β-hydroxysteroid dehydrogenase inhibitor, carbenoxolone (CBX), on CS- and DHCS-induced up-regulation of α1- and β1-mRNA in VSMC. (A) Northern blot analyses in VSMC exposed to control MEM, CS- or DHCS-supplemented MEM in the absence or presence of 10-5 m CBX. (B) Quantitations of the Northern blot. Data are expressed as a relative increase in mRNA expression compared with control samples. The number of experiments done on CS- or DHCS-treated VSMC is 5 and 6, respectively. Kidney International 1998 54, 492-508DOI: (10.1046/j.1523-1755.1998.00033.x) Copyright © 1998 International Society of Nephrology Terms and Conditions

Figure 9 Effects of CS or DHCS on Na,K-ATPase α1- and β1-subunit protein accumulation. VSMC were exposed to CS or DHCS (10-6 m) for 24hours. (A) Western blot analyses. (B) Quantitations of the Western blot. Abundance of each subunit protein is expressed as units of densitometry relative to control. Data are means ± se of 7 separate experiments. *P < 0.001 compared with control. Symbols are: (□) control; (▪) CS; (▪) DHCS. Kidney International 1998 54, 492-508DOI: (10.1046/j.1523-1755.1998.00033.x) Copyright © 1998 International Society of Nephrology Terms and Conditions

Figure 10 Effects of CS or DHCS on Na,K-ATPase activity. VSMC were treated with CS or DHCS (10-6 m) for 24 (N = 8) or 48hours (N = 7). Data are mean ± se. *P < 0.05, †P < 0.005, #P < 0.001 compared with control. Symbols are: (□) control; (▪) CS; (▪) DHCS. Kidney International 1998 54, 492-508DOI: (10.1046/j.1523-1755.1998.00033.x) Copyright © 1998 International Society of Nephrology Terms and Conditions

Figure 11 Effects of CS or DHCS on Na,K-ATPase α1- and β1-mRNA accumulation in rat aortae. Bilaterally adrenalectomyzed rats were subcutaneously treated with CS, DHCS, or vehicle for 12hours. (A) Northern blot analyses. (B) Quantitations of the Northern blot. Data are expressed as a relative increase in mRNA expression compared with control samples. The number of control, CS- or DHCS-treated rats is 5. *P < 0.001 compared with control. Kidney International 1998 54, 492-508DOI: (10.1046/j.1523-1755.1998.00033.x) Copyright © 1998 International Society of Nephrology Terms and Conditions