Chien-Te Lee, Viet M. Huynh, Li-Wen Lai, Yeong-Hau H. Lien

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Cyclosporine A-induced hypercalciuria in calbindin-D28k knockout and wild-type mice Chien-Te Lee, Viet M. Huynh, Li-Wen Lai, Yeong-Hau H. Lien Kidney International Volume 62, Issue 6, Pages 2055-2061 (December 2002) DOI: 10.1046/j.1523-1755.2002.00670.x Copyright © 2002 International Society of Nephrology Terms and Conditions

Figure 1 RT-PCR analysis of the effect of cyclosporine A (CsA) on the level of calbindin-D28k mRNA expressed in the kidney of wild-type (WT) mice. Lane 1, untreated; lane 3, CsA-treated. The cDNA product for calbindin-D28k is shown in the upper panel and β-actin, which serves as a control, in the lower panel. Calbindin-D28k was not detectable in the kidney of knockout (KO) mice (lane 2). The bar graph beneath shows that the relative calbindin-D28k/β-actin ratio was significantly reduced in CsA treated wild type mice (39 ± 4% vs. 100 ± 2%, *P < 0.01 vs. untreated WT. N = 4 in each group). Kidney International 2002 62, 2055-2061DOI: (10.1046/j.1523-1755.2002.00670.x) Copyright © 2002 International Society of Nephrology Terms and Conditions

Figure 2 Western blot analysis of the effect of cyclosporine A (CsA) on calbindin D-28k expression in wild-type (WT) mice. Lane 1, untreated; lane 3, CsA-treated; lane 4, 7 days after CsA withdrawal. Calbindin-D28k was not detectable in the kidney of knockout (KO) mice (lane 3). The bar graph beneath shows that CsA treatment for 7 days resulted in a 52% reduction in calbindin-D28k protein level (48 ± 6 vs. 100 ± 7%, *P < 0.01 vs. untreated WT, N = 4 in each group). This effect was completely reversed one week after discontinuing CsA (102 ± 9%, P > 0.05 vs. untreated WT). Kidney International 2002 62, 2055-2061DOI: (10.1046/j.1523-1755.2002.00670.x) Copyright © 2002 International Society of Nephrology Terms and Conditions

Figure 3 Cyclosporine A (CsA) does not affect the level of epithelial calcium channel (ECaC) (A), and α1G subunit of voltage-gated calcium channel (α1G) (B) mRNA expressed in the kidney of wild-type (WT) and calbindin-D28k knockout (KO) mice. Quantitative RT-PCR analysis was performed as described in the Methods section. Lane 1, untreated WT; lane 2, CsA-treated WT; lane 3, untreated KO; lane 4, CsA-treated KO mice. N = 4 in each group. Kidney International 2002 62, 2055-2061DOI: (10.1046/j.1523-1755.2002.00670.x) Copyright © 2002 International Society of Nephrology Terms and Conditions