DIV 14 neurons were transfected with siRNAs, then pretreated for 30 min with KN62 (10 μM), followed by treatment with NMDA (10 μM) and glycine (2 μM).
-
A
Samples were collected 3 min after stimulation with NMDA and glycine, and the expressions of p‐CREB, CREB, and PHF2 were examined by immunoblotting.
-
B
The mRNA levels of memory formation‐related genes were analyzed using RT–qPCR normalized to 18S rRNA levels.
-
C–E
ChIP analysis represented the binding of PHF2 (C), CREB (D), and H3K9me2 (E) to Bdnf and Nr2b promoters in neurons.
-
F
ChIP‐ChIP analysis showed that PHF2 and CREB are recruited together to the same region of Bdnf and Nr2b promoters.
Data information: In (A–F), data are representative of three independent experiments. In (B–F), data are presented as the mean ± SD (
n = 3). ***
P < 0.001, **
P < 0.01, and *
P < 0.05 (unpaired, two‐sided Student's
t‐test).
Source data are available online for this figure.