Effect of NRF-2 silencing and over-expression, with and without KCl or
TTX treatment, respectively, on the transcript levels of the AMPA receptor
subunit genes in visual cortical neurons. (A) NRF-2α shRNA transfection
in primary neurons down-regulated mRNA levels of NRF-2α
and Gria2, but not those of Gria1,
Gria3, and Gria4. Primary neurons treated
for 5 h with 20 mM KCl revealed an up-regulation of all transcripts as compared
to pBS/U6 empty vector controls. In the presence of shRNA against
NRF-2α, 5 h treatment with 20 mM KCl did not up-regulate transcripts of
NRF-2α and Gria2, but it did up-regulate
those of Gria1, Gria3, and Gria4. N = 3 for each
data point. ***= P < 0.001, **= P
< 0.01 and *= P < 0.05 when compared to pBS/U6
empty vector controls. ### = P
< 0.001 and X = non-significant when compared to KCl- treated samples.
(B) In primary neurons, NRF-2α/β over-expression led to an
increase in the transcript levels of NRF2α and Gria2,
but not those of Gria1, Gria3, and
Gria4. Primary neurons treated for 3 days with 0.4
μM TTX revealed a down-regulation of all tested transcripts as compared
to pcDNA3.1 empty vector controls. Over-expression of NRF-2α and
β rescued the down-regulation of Gria2 transcripts, but
not those of Gria1, Gria3, and Gria4. N = 3
for each data point; ***= P < 0.001 and **=
P < 0.01 when compared to controls;
### = P < 0.001 and X =
non-significant when compared to TTX-treated samples.