Induction of lysine acetylation in 14-3-3β and reduction of serine phosphorylation in synaptopodin in ADR nephropathy.

<p>Whole kidney lysates from normal saline treated mice or from mice 7, 14 or 28 days after ADR administration were examined by immunoprecipitation (IP)/Western blotting (WB) analysis. (A) IP for 14-3-3β followed by (top blot) WB for acetylated lysine showing a progressive increase in 14-3-3β acetylation in ADR nephropathy, or (middle blot) WB for synaptopodin showing binding between 14-3-3β and synaptopodin, and (bottom blot) WB for 14-3-3β showing equivalent IP of 14-3-3β from all lysates. (B) IP for synaptopodin followed by (top blot) WB for phosphor-serine showing a progressive loss of synaptopodin phosphorylation in ADR nephropathy, or (middle blot) WB for synaptopodin showing a progressive loss of synaptopodin expression in the progression of ADR nephropathy; (bottom blot) lysates were probed for α-tubulin to confirm equivalent protein content. (C) Graph shows densitometric analysis of the ratio of acetylated lysine to total 14-3-3β. Data are mean ± SD, n = 6. <i>^*P&lt;0.05 vs</i> NS 28d; <i>^#P&lt;0.05</i> vs ADR 7d. (D) Graphs show densitometric analysis of the ratio of phospho-serine to total synaptopodin, the ratio of phospho-serine to α-tubulin, and the ratio of synaptopodin to α-tubulin (D). Data are mean ± SD, n = 6. <i>^*P&lt;0.05 vs</i> NS 28d; <i>^#P&lt;0.05 vs</i> ADR 7d; ^$<i>P&lt;0.05 vs</i> ADR 14d.