Figure 3. Phosphorylation of EB2 Ser222 is suppressed by NMDAR activity.

• A–C
  Protein lysates were obtained from cortical neuronal cultures treated with 50 mM KCl applied directly to culture media for indicated durations. 100 μm AP5 was used to block NMDAR activity during KCl treatment. Quantification of phosphorylated EB2 (B) and total CDKL5 (C) is normalized to 0 min time point. EB2 phosphorylation is significantly decreased after 10 min of KCl treatment compared to 0 min. CDKL5 protein levels are slightly reduced after 40 min compared to 0 min. Dunnett's multiple comparison: n = 3 replicates.
• D, E
  Protein lysates were obtained from cortical neuronal cultures treated with 50 μM NMDA applied directly to culture media for indicated durations. Quantification of phosphorylated EB2 and total CDKL5 is normalized to 0 min time point. EB2 S222 phosphorylation is significantly reduced after 10 min compared to 0 min. Dunnett's multiple comparison: n = 3 replicates.

Data information: *P < 0.05, **P < 0.01, ***P < 0.001, error bars are SEM.Source data are available online for this figure.