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. 2016 Aug 4;6:31070. doi: 10.1038/srep31070

Figure 5. The role of PACSIN1 in regulating NMDA-induced AMPAR endocytosis is independent of PICK1.

Figure 5

(a) Cultured hippocampal neurons were transfected with empty pSuper vector (control), pSuper-PICK1-shRNA (PICK1 KD), pSuper-PACSIN1-shRNA#1 (PACSIN1 KD) or a combination of PICK1 and PACSIN1 shRNA constructs (double KD) together with the myc-GluA2 reporter at DIV15. At DIV17, neurons were stimulated with 50 μM NMDA for 5 min and the extent of myc-GluA2 internalization was monitored 5 min (t = 10 min) and 15 min (t = 20 min) after stimulation. Changes in pH-GluA2 fluorescence intensity were monitored by live-cell confocal microscopy (scale bar, 50 μm). Quantification of myc-GluA2 internalization at different time points (b), at t = 10 min (c) and at t = 20 min (d). Data represent mean ± s.e.m. (One-way ANOVA, *P < 0.05, **P < 0.01 against control cells, #P < 0.05 against PICK1 KD cells, n = 8 (Control), 6 (PICK1 KD), 6 (PACSIN1 KD), 6 (double KD) neurons for each time point from 2 independent cultures).