Western blot analysis for ACE2 protein expression in urine and ACE2, ADAM17 and TIMP3 protein expression in the kidney of <i>db/db</i>, <i>db/db</i>+rosiglitazone mice.

<p>(A) Western blot analysis for ACE2 protein expression in urine. Lane 1 represents mouse kidney lysate (positive control), lane 2 and 3 represent urine samples from 6 wks old control mice, lanes 4 and 5 represent urine samples from 6 wks old <i>db/db</i> mice, lanes 6 and 7 represent urine samples from 7 wks old <i>db/db</i>+rosiglitazone mice. Immunoreactive band for ACE2 was observed at 90 kDa in mouse kidney lysate (lane 1), but was seen at ∼70 kDa in the urine (lanes 4 and 5). One-way ANOVA showed that urinary ACE2 protein excretion significantly increased in <i>db/db</i> mice compared to their age-matched lean control mice. Rosiglitazone treatment decreased urinary ACE2 protein excretion compared to untreated <i>db/db</i> mice. *<i>p</i><0.01 Vs age-matched lean control mice. ^#<i>p</i><0.05 Vs untreated <i>db/db</i> mice. Each bar represents mean ± SEM of group size (n = 6–8). (B) Western blot analysis for ACE2 protein expression in kidney. One-way ANOVA showed that ACE2 protein expression was significantly increased in <i>db/db</i> mice compared to their age-matched lean control mice. There was no significant difference in the ACE2 protein expression in the treated <i>db/db</i> mice compared to untreated <i>db/db</i> mice after eight weeks of treatment. Each bar represents mean ± SEM of group size (n = 4–5). (C) Western blot analysis for renal ADAM17 protein expression. One-way ANOVA showed that renal ADAM17 significantly increased in <i>db/db</i> mice compared to their age-matched lean control mice. ADAM17 protein expression was significantly decreased in treated <i>db/db</i> mice compared to untreated <i>db/db</i> mice after eight weeks of treatment. *<i>p</i><0.05 Vs age-matched lean control mice. Each bar represents mean ± SEM of group size (n = 3–5). (D) Western blot analysis for renal TIMP3 protein expression. One-way ANOVA showed that renal TIMP3 significantly decreased in untreated and treated <i>db/db</i> mice compared to 30 wks old lean control mice. *<i>p</i><0.01 Vs lean control mice. Each bar represents mean ± SEM of group size (n = 6).