A. Relative CAT activity in E18 cortical neurons transfected with mouse -350 c-fos-CAT reporter along with shRNA against GFP and rat BRG1 at 3DIV, and stimulated as indicated at 5 DIV (KCl 50 mM).
B. Relative CAT activity in rat E18 cortical neurons transfected with mouse -350 c-fos-CAT reporter along with b-galactosidase and wild type BRG1 constructs at 3DIV, and stimulated as indicated at 5 DIV (KCl 50 mM) with the presence of DMSO.
C. Relative CAT activity in rat E18 cortical neurons transfected with mouse -350 c-fos-CAT reporter along with b-galactosidase and wild type BRG1 construct at 3DIV, and stimulated as indicated at 5 DIV (KCl 50 mM) with TSA pre-treatment (TSA 250ng/ml).
D. Immunoprecipitation of BRG1 with class I HDAC complex. Cortices from postnatal 4 day ratx were dissected, homogenized and immunoprecipitated with antibodies as indicated. The immunoprecipitates were resolved by SDS-PAGE and the Western blot was probed with an anti-BRG1 antibody.
E. Immunoprecipitation of BRG1 with Rb. Nuclear fractions of rat E18 cortical neurons were collected at 5DIV, and immunoprecipitated with IgG and anti-Rb antibody. The immunoprecipitates were resolved by SDS-PAGE and Western blot was probed with an anti-BRG1 antibody.
F. Immunoprecipitation of Rb with HDAC1. Nuclear fractions of rat E18 cortical neurons were collected at 5DIV, and immunoprecipitated with IgG and anti-HDAC1 antibody. The immunoprecipitates were resolved by SDS-PAGE and Western blot was probed with an anti-Rb antibody.
G. Relative CAT activity in rat E18 cortical neurons transfected with mouse -350 c-fos-CAT reporter along with b-galactosidase, wild type BRG1 and mutated BRG1 (LTCEERTREE) constructs at 3DIV, and stimulated as indicated at 5 DIV (KCl 50 mM).
H. Relative CAT activity in rat E18 cortical neurons transfected with mouse -350 c-fos-CAT reporter along with b-galactosidase, wild type Retinoblastoma protein and mutated Rb protein (Δexon 22) constructs at 3DIV, and stimulated as indicated at 5 DIV (KCl 50 mM).
Asterisks indicate significance at p<0.05. Error bars represent +SD.