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. 2014 Sep 10;8:280. doi: 10.3389/fncel.2014.00280

FIGURE 6.

FIGURE 6

T-brain-1 upregulation requires activation of CaMKII in neuronal cultures. Cultured cortical neurons of WT mice were pretreated with APV (100 μM), NBQX (100 μM), KN-93 (5 μM), or CsA (5 μM) for 30 min prior to bicuculline stimulation (Bicu, 40 μM) for 6 h. Neurons were then fixed for immunostaining (A,B) or harvested for quantitative-PCR (C) at 21 DIV. (A) Double immunostaining with TBR1 and MAP2 antibodies. Scale bars: 20 μm. The quantitative data of TBR1 fluorescence intensities are shown in (B). The number of neurons (n) for each experiment is indicated in the figures. All data represent the mean + SEM. (C) Real-time PCR to quantify the relative mRNA levels of Tbr1 and Grin2b upon bicuculline activation. The Cyp expression level was used as an internal control. Experiments were repeated at least three independent times. All data represent the mean ± SEM. *P < 0.05; **P < 0.001; ***P < 0.001; n.s., not significant.