FIG. 5.
Rictor/mTORC2 is activated in type 1 diabetes. OVE26 mice (17 weeks old) were treated with phosphorothioated sense (S) or AS oligonucleotides for Rictor (90 ng·g body wt−1·day−1), administered subcutaneously by an ALZET osmotic minipump for 1 month. Mice in the control groups received saline vehicle also administered subcutaneously by an ALZET osmotic minipump. Glomeruli were isolated from the kidneys of the four groups of mice (n = 5): group 1, FVB control mice; group 2, OVE26 mice; and groups 3 and 4, OVE26 mice treated with either S or AS. (A) Relative mRNA levels of Rictor. (B) Relative mRNA levels of Akt1. (C) Relative mRNA levels of Akt2. (D) Representative Western blot of Rictor, phospho-Ser473 Akt, phospho-Thr308 Akt, phospho-Thr346 NDRG1, Akt1, Akt2, and β-actin. β-actin was included as a control for loading and the specificity of change in protein expression. (E) Histograms showing quantitation of Rictor/β-actin from five different mice in each group. (F) Histograms showing quantitation of p-Akt (Ser473)/β-actin results from five different mice in each group. (G) Histograms showing quantitation of p-Akt (Thr308)/β-actin results from five different mice in each group. (H) Histograms showing quantitation of p-NDRG1 (Thr346)/β-actin results from five different mice in each group. (I) Histograms showing quantitation of Akt1/β-actin results from five different mice in each group. (J) Histograms showing quantitation of Akt2/β-actin results from five different mice in each group. (K) Representative Western blot of Raptor, phospho-Thr389 p70S6K, p70S6K, and β-actin. (L) Histograms showing quantitation of Raptor/β-actin results from five different mice in each group. (M) Histograms showing quantitation of p-p70S6KThr389/p70S6K results from five different mice in each group. Values are mean ± SE of five different animals from each group. *p < 0.05 versus vehicle-treated FVB mice; #p < 0.05 versus vehicle-treated OVE26 mice. AS, antisense.