Increased cell migration of PC12-SH2B1β cells during wound healing associates with elevated pPLCγ1 and pPKC.

<p>PC12-GFP and PC12-SH2B1β cells were differentiated and subjected to wound healing as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0034999#pone-0034999-g001" target="_blank">Figure 1</a>. (A) Movement of cells around the wounded gaps was tracked as described in the <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0034999#s2" target="_blank">Materials and Methods</a> during healing for 6 days. The color lines represent the migration path of cells from time 0 (blue) to 144 h (red) during healing. (B) The accumulated distances and motility during time 0–144 h were calculated. Cell motility (rate of cell migration) is defined as accumulated distance/time. Values are mean ± S.E.M. from four independent experiments. (*: P<0.05, paired Student's t-test) (C) Equal amount of proteins from the harvested cell lysate of un-wounded (U) or wounded cells was resolved via SDS-PAGE and immunoblotted with anti-pPLCγ1 and anti-PLCγ1 antibodies. (D) Relative levels of pPLCγ1 were normalized to total PLCγ1 and then the levels in PC12-GFP cells on differentiated day 8 (U). Values are mean ± S.E.M. from at least three independent experiments. (*: P<0.05, one-way ANOVA) (E) Lysate or immunoprecipitated PKC were resolved via SDS-PAGE and immunoblotted with anti-PKC, pSer or pThr antibody. Relative pSer or pThr levels were normalized to immunoprecipitated PKC and then the levels in PC12-GFP cells on differentiated day 8 (un-wounded). Values are mean ± S.E.M. from three independent experiments for relative pSer levels and mean ± S.D. from two independent experiments for relative pThr levels.