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. 2015 Mar 16;112(13):4128–4133. doi: 10.1073/pnas.1422448112

Fig. 2.

Fig. 2.

Adult hippocampal NSPCs synthesize and secrete large quantities of VEGF. (A) Isolated adult hippocampal NSPCs treated with different growth factor conditions for 4 d expressed mRNA transcripts for the VEGF120 and VEGF164 but not VEGF188 splice variants. NSPCs also expressed VEGFR2 mRNA but not VEGFR1 or soluble Flt (sFlt), the secreted form of VEGFR1. Adult lung RNA served as a positive control. (B) NSPC secretion of VEGF was measured by ELISA of culture supernatant after 4 d and was greatest in standard proliferative conditions with 20 ng/mL EGF and 20 ng/mL FGF2. Decreasing either EGF or FGF2 reduced secretion of VEGF (ANOVA, P < 0.0001; n = 4–5 wells per group per experiment; two experiments). (C) Adult NSPCs were maintained in 20 ng/mL EGF and 20 ng/mL FGF2 for 2 d (0 d differentiation) or switched to differentiating conditions for 2, 4, or 8 d. VEGF ELISA showed decreased secreted VEGF protein with differentiation (ANOVA, P < 0.0001; n = 3 wells per group per experiment; three experiments). (D and E) VEGF120 and VEGF164 mRNA decreased with differentiation normalized to the MAPK3 housekeeping gene relative to 0 d differentiation (VEGF120: ANOVA, P = 0.0002; VEGF164: ANOVA, P < 0.0001; n = 2–3 wells per group per experiment; three experiments). Data represent mean ± SEM. **P < 0.01; ***P < 0.001; ****P < 0.0001, post hoc Dunnett’s tests.