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. 2013 Jan 9;33(2):790–803. doi: 10.1523/JNEUROSCI.1238-12.2013

Figure 5.

Figure 5.

Phosphorylation of Pak1, LIMK, and cofilin as well as levels in lipid rafts of the Pak1 activators PIX and cdc42 are abnormal in NCAM−/− versus NCAM+/+ growth cones. Brain homogenates (lanes 1 and 2), growth cones (lanes 3 and 4), nongrowth cone membranes (non-GC; lanes 5 and 6), and lipid rafts from growth cones (GC LR; lanes 7 and 8) and nongrowth cone membranes (non-GC LR; lanes 9 and 10) probed by Western blot with indicated antibodies. Labeling for contactin-1 and GAPDH was included as loading control. Note that lipid raft marker contactin-1 is highly enriched in lipid rafts (lanes 7–10). Levels of Thr212-phosphorylated Pak1 and cdc42 are increased in lipid rafts from NCAM−/− versus NCAM+/+ growth cones (compare lanes 7 and 8). Levels of total LIMK and cofilin are increased in NCAM−/− versus NCAM+/+ growth cones (compare lanes 3 and 4) and growth cone lipid rafts (compare lanes 7 and 8), and levels of Thr508 phosphorylated LIMK and Ser3 phosphorylated cofilin are reduced in NCAM−/− versus NCAM+/+ growth cones (compare lanes 3 and 4). Experiments and Western blot analyses were performed three times with the same results.