(a) Representative images of surface GluA1 staining of neurons transfected with a control plasmid, Arc-mNES or Arc-NES and treated with bicuculline for 48 h.
(b) Quantification of surface GluA1 staining in neurons transfected with a control plasmid, Arc-mNES or Arc-NES shows Arc localization to the nucleus prevents downscaling. N=65 for control, N=64 for bicuculline, N=37 for Arc-mNES, N=50 for Arc-mNES+bicuculline, N=50 for Arc-NES, N=50 for Arc-NES+bicuculline from four independent cultures.
(c) Quantification of Arc levels in untransfected neurons using Arc staining and neurons transfected with ABCDEFG-Arc-His-Flag-HI using Flag staining, with and without 8-h BDNF treatment. N=55 for endogenous Arc, N=37 for endogenous Arc + BDNF, N=40 for ABCDEFG-Arc-His-Flag-HI, N=37 for ABCDEFG-Arc-His-Flag-HI+BDNF from three independent cultures.
(d) Representative images of ABCDEFG-Arc-His-Flag-HI construct expression with and without BDNF treatment in neurons stained with anti-Flag antibody.
(e) Representative images of surface GluA1 staining of Arc−/− neurons transfected with a control plasmid, ABCDEFG-Arc-mNES-HI or ABCDEFG-Arc-NES-HI and treated with bicuculline for 48 h.
(f) Quantification of surface GluA1 staining in neurons transfected with a control plasmid, ABCDEFG-Arc-mNES or ABCDEFG-Arc-NES shows Arc localization to the nucleus rescues downscaling in Arc knockout neurons. N=58 for control, N=47 for control+bicuculline, N=59 for ABCDEFG-Arc-mNES-HI, N=45 for ABCDEFG-Arc-mNES-HI+bicuculline, N=40 for ABCDEFG-Arc-NES-HI, N=30 for ABCDEFG-Arc-NES-HI+bicuculline from at least three independent cultures.
***, p<0.001, n.s., not significant, by ANOVA. Error bars show 95% confidence intervals. Scale bars=10 μm.