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. 2020 Jul 3;295(35):12330–12342. doi: 10.1074/jbc.RA120.014303

Figure 3.

Figure 3.

Application of okadaic acid increases phosphorylation of GABAAR α2 Ser-359 in pHα2 mice. A, horizontal brain slices from pHα2 mice were treated with OA (1 μm) or DMSO vehicle control for 30 min. These brain lysates were then used to immunoprecipitate pHα2 and immunoblot for phosphory-lated α2 at Ser-359 (α2 pS359). Blots were incubated with α2 pS359 antibody alone (No peptide), or α2 pS359 antibody incubated with synthetic phosphorylated Ser-359 peptide (Phosphopeptide) or nonphosphorylated Ser-359 peptide (Non-phosphopeptide). Data are means ± S.E. (error bars). ****, p < 0.0001; one-way ANOVA, Tukey's post hoc test, n = 3. B, rat cortical neurons were infected at 15 DIV with lentivirus expressing pHα2 WT or the S359A mutant. At 19 DIV, neurons were treated with OA (1 μm) or DMSO vehicle control for 30 min. Cell lysates were subjected to immunoprecipitation with GFP-Trap, and immunoblots were incubated with α2 or α2 pSer-359 antibody. Data are means ± S.E. ****, p < 0.0001; one-way ANOVA, Tukey's post hoc test, n = 4.