Figure 6

Impairment of neuronal and oligodendrocyte differentiation in PLCγ1-depleted adult NSCs. (A) A schematic diagram illustrating the retroviral shRNA construct and western blot analysis on the efficacy of knockdown by shRNAs against endogenous PLCγ1 in adult NSCs. (B) Immunostaining of endogenous PLCγ1 in NSCs infected with shRNA-i1. Scale bar: 20 μm. (C) Western blot analysis of neuronal differentiation marker Tuj1 and glial marker GFAP in PLCγ-depleted NSCs in the presence or absence of FGF-2, or after induction of differentiation with RA (0.5 μM) and FBS (0.5%) for 6 days. (D) Quantification of various cell types in PLCγ1-depleted NSCs after induction of differentiation with RA (0.5 μM) and FBS (0.5%) for 6 days. Values represent mean ± SEM. (n = 4; *: P < 0.01, Student's t-test). (E) Rescue of neuronal differentiation defects of endogenous PLCγ1-depleted adult NSCs by exogenous expression of PLCγ1 cDNA plasmid. (F) Western blot analysis of neuronal differentiation marker Tuj1 after differentiation of adult NSCs expressing WT and two different dominant negative mutants of PLCγ1 (LIM: lipase inactive mutant, SH3: SH3 domain deletion mutant).