Fig. 3

Prolonged alterations of hippocampal network activity regulate Tyr-phosphorylation and levels of GluN2B and GluA2. Immunoblot analysis of hippocampal cultured neurons that were treated for 48 h with CTL, 24–48 h TTX (a–c), or 24–48 h BC treatment (d–f) (n = 6 per treatment). a–c Prolonged TTX treatment increased the level of Tyr¹⁴⁷²–phosphorylated GluN2B (GluN2B-pY¹⁴⁷²) (a) and the level of GluA2 that were phosphorylated at Tyr ⁸⁶⁹, Tyr ⁸⁷³, and Tyr ⁸⁷⁶ (3Tyr) (GluA2-p3Y) (b). d–f Prolonged BC treatment decreased the levels of GluN2B-pY¹⁴⁷² (d) and GluA2-p3Y (e). Total GluN2B and GluA2 levels were reduced at 48 h BC application. c, f The relative phosphorylation state of GluN2B and GluA2 as calculated by the ratio of phosphorylated proteins over total proteins followed by normalization to GAPDH. Data shown represent the mean ± SEM (*p < 0.05; **p < 0.01)