Figure 1.

VEGFR inhibition in control and diabetic wildtype and eNOS^−/− mice. (A) Effect of vatalanib in cultured human renal glomerular endothelial cells. Cells were preincubated with either vatalanib (1 µM) or DMSO for 2 hours before stimulation with 20 ng/ml VEGF for 5 minutes. Cell lysates were immunoprecipitated for VEGFR-2 and immunoblotted for either phosphotyrosine or VEGFR-2. (B) SBP in C57BL/6 mice treated with vehicle or vatalanib (10 mg/kg, 50 mg/kg, 100 mg/kg daily) for 7 days. (C) pVEGFR-2 and total VEGFR-2 in kidney homogenates from control and STZ-diabetic wild-type (C57BL/6) and eNOS^−/− mice treated with vehicle or vatalanib (50 mg/kg) for 12 weeks (n=3/group). (D–K) Glomerular PECAM-1 staining in mice treated with vehicle or vatalanib (50 mg/kg) for 12 weeks: (D) C57BL6 + vehicle, (E) C57BL/6 + vatalanib, (F) STZ-C57BL/6 + vehicle, (G) STZ-C57BL/6 + vatalanib, (H) eNOS^−/− + vehicle, (I) eNOS^−/− + vatalanib, (J) STZ-eNOS^−/− + vehicle, and (K) STZ-eNOS^−/− + vatalanib. Quantitation of glomerular PECAM-1 immunostaining in (L) C57BL/6 mice and (M) eNOS^−/− mice. (N–U) Glomerular capillary volume determined by FMA in mice treated with vehicle or vatalanib (50 mg/kg) for 12 weeks: (N) C57BL6 + vehicle, (O) C57BL/6 + vatalanib, (P) STZ-C57BL/6 + vehicle, (Q) STZ-C57BL/6 + vatalanib, (R) eNOS^−/− + vehicle, (S) eNOS^−/− + vatalanib, (T) STZ-eNOS^−/− + vehicle, and (U) STZ-eNOS^−/− + vatalanib. Quantitation of glomerular capillary volume in (V) C57BL/6 and (W) eNOS^−/− mice. *P<0.05 versus 0 mg/kg per day, ^†P<0.05 versus all other groups, ^‡P<0.01 versus all other groups, ^§P<0.05 versus eNOS^−/− + vehicle. AU, arbitrary units. Scale bar, 30 µm. Original magnification, ×400.