Volume 74, Issue 4, Pages (August 2008)

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Volume 74, Issue 4, Pages 478-485 (August 2008) PGE2 inhibits basolateral 50pS potassium channels in the thick ascending limb of the rat kidney Ruimin Gu, Yan Jin, Yuanyuan Zhai, Lei Yang, Chengbiao Zhang, Wennan Li, lijun Wang, Shumin Kong, Baofeng Yang, Wen-Hui Wang Kidney International Volume 74, Issue 4, Pages 478-485 (August 2008) DOI: 10.1038/ki.2008.198 Copyright © 2008 International Society of Nephrology Terms and Conditions

Figure 1 A channel recording demonstrating the effect of 10μM PGE2 on the basolateral 50pS K channels. The experiment was performed on a cell-attached patch and the holding potential was −20mV (depolarization). The channel close level is indicated by a dotted line and C. Two parts of the record indicated by numbers are extended to show the fast time resolution. Kidney International 2008 74, 478-485DOI: (10.1038/ki.2008.198) Copyright © 2008 International Society of Nephrology Terms and Conditions

Figure 2 A channel recording showing the effect of 10μM sulprostone on the basolateral 50pS K channels. The experiment was performed on a cell-attached patch and the holding potential was 0mV. The channel close level is indicated by a dotted line and C. Two parts of the record indicated by numbers are extended to show the fast time resolution. Kidney International 2008 74, 478-485DOI: (10.1038/ki.2008.198) Copyright © 2008 International Society of Nephrology Terms and Conditions

Figure 3 A channel recording showing the effect of calphostin-C on the 50pS K channels in the TAL. The experiment was performed on a cell-attached patch and the holding potential was −20mV (depolarization). The channel close level is indicated by a dotted line and C. Two parts of the record indicated by numbers are extended to show the fast time resolution. Kidney International 2008 74, 478-485DOI: (10.1038/ki.2008.198) Copyright © 2008 International Society of Nephrology Terms and Conditions

Figure 4 A channel recording demonstrating the effect of 10μM PGE2 on the basolateral 50pS K channels in the absence and presence of 300nM calphostin-C. The experiment was performed on a cell-attached patch and the holding potential was 0mV. The channel close level is indicated by a dotted line and C. Three parts of the record indicated by numbers are extended to show the fast time resolution. Kidney International 2008 74, 478-485DOI: (10.1038/ki.2008.198) Copyright © 2008 International Society of Nephrology Terms and Conditions

Figure 5 Effect of PGE2 (10μM), PGE2+calphostin-C (cal.c) (300nM), and phorbol 12-myristate 13-acetate (5μM) on the basolateral 50pS K channels. The experiments were carried out on cell-attached patches. Kidney International 2008 74, 478-485DOI: (10.1038/ki.2008.198) Copyright © 2008 International Society of Nephrology Terms and Conditions

Figure 6 A channel recording demonstrating the effect of PD98059 (50μM) and 10μM PGE2+PD98059 on the basolateral 50pS K channels in the TAL. The experiment was performed on a cell-attached patch and the holding potential was −10mV (depolarization). The channel close level is indicated by a dotted line and C. Three parts of the record indicated by numbers are extended to show the fast time resolution. Kidney International 2008 74, 478-485DOI: (10.1038/ki.2008.198) Copyright © 2008 International Society of Nephrology Terms and Conditions

Figure 7 Effect of 10μM PGE2 on the basolateral 50pS K channels in the TAL treated with PD98059, SB203580 or both agents. The experiments were performed on cell-attached patches. The asterisk indicates the significant difference between the MAPK inhibitor and the corresponding control. # Indicates a significant difference between the PD group and PD+PGE2 group or between the SB and SB+PGE2 group. Kidney International 2008 74, 478-485DOI: (10.1038/ki.2008.198) Copyright © 2008 International Society of Nephrology Terms and Conditions

Figure 8 Effect of PGE2 on the phosphorylation of p38 and ERK. Western blots show the effect of 10μM PGE2 on the phosphorylation of p38 (a) and ERK (b) in the presence or absence of 10μM indomethacin. Data are summarized in a bar graph (bottom panel). The asterisk indicates a significant difference in comparison with the control. Kidney International 2008 74, 478-485DOI: (10.1038/ki.2008.198) Copyright © 2008 International Society of Nephrology Terms and Conditions

Figure 9 Effect of PGE2 on the phosphorylation of p38 and ERK in the presence of PKC inhibitor. Western blots show the effect of PGE2 on the phosphorylation of p38 (a) and ERK (b) in the presence or absence of 300nM calphostin-C. Data are summarized in a bar graph (bottom panel). The asterisk indicates a significant difference in comparison with the control. Kidney International 2008 74, 478-485DOI: (10.1038/ki.2008.198) Copyright © 2008 International Society of Nephrology Terms and Conditions

Figure 10 A cell model illustrating the mechanism by which PGE2 inhibits the basolateral 50pS K channels by a PKC–MAPK pathway in the TAL. Kidney International 2008 74, 478-485DOI: (10.1038/ki.2008.198) Copyright © 2008 International Society of Nephrology Terms and Conditions