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. Author manuscript; available in PMC: 2013 Apr 1.
Published in final edited form as: Kidney Int. 2011 Jan 12;79(8):883–896. doi: 10.1038/ki.2010.526

Figure 6. Effect of RBX on IRS1 and NO synthesis in the glomeruli of diabetic SD rats and ZF rats.

Figure 6

A and B. Representative immunoblots of tyrosine phosphorylation of IRS1 from glomerular and tubular fractions. Solubilized glomeruli and tubular fractions were isolated and subjected to immunoprecipitation followed by immunoblotting. Data from three experiments were quantitated by densitometry. A) Cont. vs. Cont.+RBX vs. DM vs. DM+RBX, n=6 in Cont., Cont.+RBX and DM. n=5 in DM+RBX, **P<0.001 vs. Cont./insulin(-)/RBX(-). ##P<0.001 vs. Cont./insulin(+)/RBX(-). ††P<0.001 vs. DM/insulin(+)/RBX(-). P<0.001 vs. DM/insulin(-)/RBX(-). B) ZL vs. ZL+RBX vs. ZF vs. ZF+RBX n=6 in each group, **P<0.001 vs. ZL/insulin(-)/RBX(-). ##P<0.001 vs. ZL/insulin(+)/RBX(-). ††P<0.001 vs. ZF/insulin(+)/RBX(-).

C. Solubilized glomeruli fractions were immunoprecipitated with antibodies against ubiquitin and subsequently immunoblotted with anti-IRS1 antibodies. Data from three experiments were quantitated by densitometry, n=6 in Cont., Cont.+RBX and DM. n=5 in DM+RBX, **P<0.001 vs. Cont./RBX(-), ††P<0.001 vs. DM/RBX(-).

D and E. The effect of RBX on NO synthesis in the glomeruli of diabetic SD rats and ZF rats. Isolated glomeruli from each group were incubated with insulin (100nM for 30 minutes). After homogenized and centrifuged, supernatant were collected. NO levels in the supernatant were measured with the Nitric Oxide Colorimeric Assay Kit. The results were derived from three separate experiments. D) **P<0.001 vs. Cont./insulin(-)/RBX(-). ##P<0.001 vs. Cont./insulin(+)/RBX(-). †P<0.05 vs. DM/insulin(+)/RBX(-). E) **P<0.001 vs. ZL/insulin(-)/RBX(-). ##P<0.001 vs. ZL/insulin(+)/RBX(-). †P<0.05 vs. ZF/insulin(+)/RBX(-).These data are expressed as means ± SD. Cont.=control rats, Cont.+RBX=control rats treated with ruboxistaurin, DM=STZ-induced diabetic rats, DM+RBX=STZ-induced diabetic rats treated with ruboxistaurin, ZL=Zucker lean rats, ZL+RBX= Zucker lean rats treated with ruboxistaurin, ZF=Zucker fatty rats, ZF+RBX= Zucker fatty rats treated with ruboxistaurin.

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