Volume 53, Issue 4, Pages 926-931 (April 1998) Subtractive hybridization cloning: An efficient technique to detect overexpressed mRNAs in diabetic nephropathy Technical Note  Marie-Noëlle Peraldi, M.D., Jeannig Berrou, Jacqueline Hagège, Eric Rondeau, Jean-Daniel Sraer  Kidney International  Volume 53, Issue 4, Pages 926-931 (April 1998) DOI: 10.1111/j.1523-1755.1998.00834.x Copyright © 1998 International Society of Nephrology Terms and Conditions

Figure 1 Schema of the method used to prepare the subtractive probe. Kidney International 1998 53, 926-931DOI: (10.1111/j.1523-1755.1998.00834.x) Copyright © 1998 International Society of Nephrology Terms and Conditions

Figure 2 Northern blot analysis. Twenty micrograms of total RNA from control (lane 1) and diabetic rat cortex kidneys after 15 days (lane 2) and 99 days (lane 3) of diabetic course. The blot was probed with 32P-α-labeled mdr 1 clone (upper panel) and alpha-actin probe (lower panel). Arrows correspond to 28 S and 18 S RNA. Kidney International 1998 53, 926-931DOI: (10.1111/j.1523-1755.1998.00834.x) Copyright © 1998 International Society of Nephrology Terms and Conditions

Figure 3 Western blot analysis. Twenty-five micrograms of control (lane 1) and diabetic (lane 2) cortex rat kidney proteins were prepared at day 99, and run on a 7.5 polyacrylamide gel. The blot was probed with the MRK 16 monoclonal antibody and revealed using the alkaline phosphatase system. Arrows correspond to 132 kDa and 84 kDa. Kidney International 1998 53, 926-931DOI: (10.1111/j.1523-1755.1998.00834.x) Copyright © 1998 International Society of Nephrology Terms and Conditions

Figure 4 Northern blot analysis. Twenty micrograms of total RNA from control (lane 1) and diabetic rat cortex kidneys after 15 days (lane 2) and 99 days (lane 3) of diabetic course. The blot was probed with 32P-α-labeled βPP clone (upper panel) and α-actin probe (lower panel). Arrows correspond to 28 S and 18 S RNA. Kidney International 1998 53, 926-931DOI: (10.1111/j.1523-1755.1998.00834.x) Copyright © 1998 International Society of Nephrology Terms and Conditions

Figure 5 Western blot analysis. Twenty-five micrograms of control (lane 1) and diabetic (lane 2) cortex rat kidney proteins were prepared at day 99, and run on a 7.5 polyacrylamide gel. The blot was probed with the Alz 90 monoclonal antibody and revealed using the chemoluminescence system. Arrows correspond to 84 kDa and 47 kDa. Kidney International 1998 53, 926-931DOI: (10.1111/j.1523-1755.1998.00834.x) Copyright © 1998 International Society of Nephrology Terms and Conditions

Figure 6 β-amyloid protein precursor (βPP) detection in rat kidney sections using the monoclonal anti-βPP 22C11 and the APAAP system. (A) Control kidney section (×500). (B) Diabetic (day 99) rat kidney section (×500). Reproduction of this figure in color was made possible by a grant from Produits Roche (Neuille sur Seine, France). Kidney International 1998 53, 926-931DOI: (10.1111/j.1523-1755.1998.00834.x) Copyright © 1998 International Society of Nephrology Terms and Conditions

Figure 7 Aβ detection in human kidneys and brain sections using the monoclonal anti-human Aβ antibody and the biotin-streptavidin-peroxydase revelation system. (A) Brain section from a patient who died of Alzheimer's disease (×250). (B) Kidney section from a patient with renal failure due to IgA nephropathy (×250). (C) Kidney section from a patient after 56 years of insulin-dependant diabetic course (×500). (D) Kidney section a patient after 17 years of insulin-dependant diabetic course; the arrows correspond to the fine Aβ deposits (×250). Reproduction of this figure in color was made possible by a grant from Produits Roche (Neuille sur Seine, France). Kidney International 1998 53, 926-931DOI: (10.1111/j.1523-1755.1998.00834.x) Copyright © 1998 International Society of Nephrology Terms and Conditions