FIG. 3.

Endogenous REST/NRSF is expressed in proliferating NSCs and the expression is repressed as the NSCs enter neuronal differentiation. (A) Endogenous mouse REST/NRSF (mREST) and transgenic human REST/NRSF (hREST) transcription was assessed by RT-PCR in NSC-M-R, NSC-M-V, and NSC-M cells growing in proliferation medium (day 0) or in differentiation medium in vitro for 2, 4, and 6 days. Actin transcripts were measured as an internal control. (B) The percentage of REST transcriptional repression activity was determined in NSC-M, NSC-M-V, and NSC-M-R cells growing in proliferation medium (day 0) or differentiation medium for 4 days (day 4). The cells were cultured either with (+D) or without (−D) doxycycline, transfected with the reporter plasmid pNaCh (+RE) or pNaCh (−RE), and analyzed as described in the text. The data represent percent REST activity.