Figure 5.

Neto2 Ser-409 phosphorylation is reduced upon co-expression with KARs. A, Neto2, CaMKII (T286D), and the indicated KAR subunit (HA-GluK1 or GluK2) were expressed in COS-7 cells and probed with Ser(P)-409 antibody. The blots were also probed with Neto2 antibody to determine total Neto2 amounts. B, mean ± S.E. (n = 5 for GluK1 and n = 3 for GluK2) of phosphorylated Neto2. Quantitation was done by densitometry, and a ratio of phosphorylated/total Neto2 is plotted (*, p < 0.05; n.s., non-significant). Error bars represent S.E. C, HA-GluK1 and Neto2 (WT or mutants) were co-expressed. The upper panel shows the immunoprecipitated (IP) GluK1 and the bound Neto2 using HA and Neto2 antibodies, respectively. Neto2 expression for different mutants was tested in the lysate alone (lower panel).