Localized inactivation of GSK-3β was associated with the formation of cell polarity.

<p>(A) Up-regulation of pSer-GSK-3 during the wound recovery of U251 cells. <i>Left</i>: representative western blot results of U251 cell lysates collected at indicated time points after scratching. <i>Right</i>: statistics of p-Ser-GSK-3 levels normalized to GSK-3 levels and compared with the value at 0hr. *<i>P</i><0.05, ** <i>P</i><0.01 versus the value of pGSK-3/GSK-3 at 0hr. (B) Location of increased pGSK-3 staining in the wound recovery of U251 cells at 0 hr or 3 hrs after scratching. Green: anti-pSer21-GSK-3α or anti-pSer9-GSK-3β; Red: rhodamine phalloidin. <i>Original magnification, 400×</i>.(C) MTOC mis-orientation in U251 caused by GSK-3 inhibitors. <i>Left</i>: representative images of MTOC in different treatments. Green: anti-α-tubulin; Red: anti-pericentrin; Blue: Hoechst 33258. <i>Original magnification, 400×</i>. <i>Right</i>: quantification of centrosome polarization as described in <i>Materials and Methods</i> in U251 with or without GSK-3β inhibitors. ** <i>P</i><0.01 versus DMSO. LiCl 20mM, SB216763, 20μM, NaCl, 20 mM. Data were mean ± SD of three independent experiments.