![Rescue of defective GluA2 recycling by Flag-WDR91, Flag-WDR91(3A), and Flag-WDR91(Δ5) in Wdr91−/−mouse primary neurons. (A) Time-course recording of pH-GluA2 in mouse primary hippocampal neurons with the indicated genotypes. Neurons were transfected with pH-GluA2– and WDR91-expressing vectors and subjected to glutamate perfusion for 5 min. Glutamate was removed at the time point 5 min, and neurons were continuously imaged. Bars, 5 μm. (B) Time-course tracing of pH-GluA2 fluorescence intensity in neurons shown in A. n = 9 neurons (Wdr91+/+); n = 7 neurons (Wdr91−/−); n = 6 neurons (Wdr91−/− + WDR91[WT]); n = 6 neurons (Wdr91−/− + WDR91[3A]); n = 6 neurons (Wdr91−/− + WDR91[Δ5]). (C) Left: Maximum amplitudes of pH-GluA2 fluorescence intensity changes upon glutamate stimulation. Right: Average half-time (T1/2) of pH-GluA2 fluorescence recovery after removal of glutamate. n = 9 neurons (Wdr91+/+); n = 7 neurons (Wdr91−/−); n = 6 neurons (Wdr91−/− + WDR91[WT]); n = 6 neurons (Wdr91−/− + WDR91[3A]); n = 6 neurons (Wdr91−/− + WDR91[Δ5]). For all quantifications, error bars represent SEM. Data are from three independent experiments. Statistical analyses were performed with the Kruskal–Wallis test. ***, P < 0.001. NS, P > 0.05.](https://cdn.rupress.org/rup/content_public/journal/jcb/221/12/10.1083_jcb.202203013/2/jcb_202203013_figs3.png?Expires=1771195935&Signature=VJniX8fuNSWdlspgyukiCqTlREUQU51g67ebS1Vf9MCEotgAM2d3cNJgFIx7eQUZAGdwf2v2bU1awtUoBvQT~juNHIWiaIW4KZgmqZxue8K2m0-wvV1E0ujj0yZsuGgAqBcp0aTPvNEp73m7NT~d9RHot9M13nfZWsBNeJ8RWGnbKbF19SX61ENmeguWO06gbIPpq5lrNaLeH1yXh-8CVGt1W5m83QWcEpnS-ywbBgjj1DI5i7NHbNJxRx6OzNLdThLZXWK8v-TrQwgpqrpArf0esY2f863W9PhVb5SMFUcS5H08AwNMDEfjy-ff7SomGQSC9zYiIKHCa0FbFws2~A__&Key-Pair-Id=APKAIE5G5CRDK6RD3PGA)
Rescue of defective GluA2 recycling by Flag-WDR91, Flag-WDR91(3A), and Flag-WDR91(Δ5) in Wdr91 −/− mouse primary neurons. (A) Time-course recording of pH-GluA2 in mouse primary hippocampal neurons with the indicated genotypes. Neurons were transfected with pH-GluA2– and WDR91-expressing vectors and subjected to glutamate perfusion for 5 min. Glutamate was removed at the time point 5 min, and neurons were continuously imaged. Bars, 5 μm. (B) Time-course tracing of pH-GluA2 fluorescence intensity in neurons shown in A. n = 9 neurons (Wdr91+/+); n = 7 neurons (Wdr91−/−); n = 6 neurons (Wdr91−/− + WDR91[WT]); n = 6 neurons (Wdr91−/− + WDR91[3A]); n = 6 neurons (Wdr91−/− + WDR91[Δ5]). (C) Left: Maximum amplitudes of pH-GluA2 fluorescence intensity changes upon glutamate stimulation. Right: Average half-time (T1/2) of pH-GluA2 fluorescence recovery after removal of glutamate. n = 9 neurons (Wdr91+/+); n = 7 neurons (Wdr91−/−); n = 6 neurons (Wdr91−/− + WDR91[WT]); n = 6 neurons (Wdr91−/− + WDR91[3A]); n = 6 neurons (Wdr91−/− + WDR91[Δ5]). For all quantifications, error bars represent SEM. Data are from three independent experiments. Statistical analyses were performed with the Kruskal–Wallis test. ***, P < 0.001. NS, P > 0.05.
