Figure 7.

PI3K activity is required for the regulation of NPC proliferation by RhoG-V12. (A) Twenty-four hours after control or RhoG-V12 vector was electroporated at E13, the regions of cortices containing EYFP⁺ cells were dissociated and dispersed on PDL-coated coverslips. Cells were cultured for 24 h in the absence or presence of LY294002 (20 μM) and labeled with BrdU for 3 h before fixation. Control, n = 6; RhoG-V12, n = 6. Values indicate means ± SEM; ***p < 0.001 (Student's t test). (B) Twenty-four hours after control or RhoG-V12 vector was electroporated at E13, the regions of cortices containing EYFP⁺ cells were dissociated and dispersed on PDL-coated coverslips. Cells were cultured for 48 h in the absense or presence of LY294002 (20 μM) and then fixed and stained with anti-Nestin antibody. Control, n = 6; RhoG-V12, n = 6. Values indicate means ± SEM; ***p < 0.001 (Student's t test). (C) NPCs derived from E12 cortices transfected with control or RhoG-V12 were analyzed by immunoblotting with antibodies against phosphorylated Akt at serine 473 (p-Akt) and Akt (Total Akt). Quantitative analysis shows the amount of phosphorylated Akt normalized to the amount of total Akt in cell lysates. Value from control cells was arbitrarily set at 1.0. Values indicate means ± SEM of three independent experiments. *p < 0.05 (Student's t test).