FIGURE 7:

PI3-K δ regulates R1881- and NGF-induced neurite outgrowth. Quiescent PC12 cells on polylysine-coated coverslips were used. (A, B) Cells were left untreated or treated for 48 h with R1881 (10 nM) or NGF (100 ng/ml) in the absence or presence of the PI3-K δ inhibitor IC87114 (5 μM). Neurite outgrowth was analyzed by contrast phase microscopy and expressed as percentage of total cells. Means and SEM are shown. The statistical significance of results was also evaluated by paired t test. The difference in neurite outgrowth was significant (*p < 0.05) only between cells challenged with R1881 or NGF and unstimulated cells. (B) Images from one experiment in A. Plastic-plated quiescent PC12 cells were used. Cells were left untreated or treated for 5 min with 10 nM R1881 (C) or 100 ng/ml NGF (D) in the absence or presence of the indicated compounds. Bicalutamide (Bic) was used at 10 μM; S peptide was used at 1 nM; IC87114 (IC) and EHT1864 were both used at 5 μM. Akt activation (P-Akt) was analyzed in lysate proteins using the anti–P-Ser-473 Akt Ab. Filters were reprobed with anti-Akt (Akt) Ab.