Fig. 4.
Pharmacological glycogen synthase kinase-3β (GSK-3β) inhibition decreases phosphorylated nucleophosmin (pNPM) threonine 95 (T95) accumulation in human and murine primary renal cells and kidney cortical homogenates during acute stress. A: immunoblot analysis of steady-state pNPM T95 content in the cytosolic fraction of primary human (h) and murine (m) proximal tubule epithelial cells (PTECs) after metabolic stress (ATP depletion × 70 min) in the presence of 4-benzyl-2-methyl-1,2,4-thiadiazolidine-3,5-dione (TDZD-8; 2 or 10 μM, 6-h pretreatment) or vehicle control (−). GAPDH was used as a loading control. B: TDZD-8 (4 mg/kg body wt ip 2 h before ischemia) decreased NPM T95 phosphorylation in the cytosolic fraction of renal cortical homogenates harvested from 8-wk-old male and female mice after ischemia in vivo.