Figure 5.

Ca²⁺/CaM-dependent protein kinase activity is required for mGluR1/5 signaling in primary hippocampal neurons. DIV 8 mouse hippocampal neurons were pre-treated with 10 uM KN62 (A to C), 5 uM KN93 (D to F), or vehicle control for 30 min, after which the cells were treated with 100 uM DHPG or vehicle for 30 min. The levels of pERK1/2 (A, D), pS6K1 (at Thr421/Ser424) (B, E), and Arc (C, F) were determined by Western blot. Representative images are shown in the upper panels and quantifications are shown in the lower panels. The level of pERK1/2 and pS6K1 was normalized to the level of total ERK1/2 and total S6K1, respectively. The level of Arc was normalized to the level of β-actin. The relative protein level in the control group was defined as 1, and all samples were normalized to the control group. All data were collected from 6 independent samples. *: p < 0.05 between control and the DHPG-treated samples. #: p < 0.05 between the indicated group and the DHPG-treated group. The p value was determined by one-way ANOVA and post hoc LSD test.