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Fig. 6 | Molecular Brain

Fig. 6

From: Regulated internalization of NMDA receptors drives PKD1-mediated suppression of the activity of residual cell-surface NMDA receptors

Fig. 6

Knockdown of PKD1 does not affect NMDAR internalization but prevents the NMDAR internalization-induced increases in serine phosphorylation of surface NMDARs. a - c DHPG or high NMDA/glycine (N + G) induced NMDAR internalization in neurons infected with PKD1 shRNA. After biotinylation of these neurons, GluN1 (a), GluN2A (b) or GluN2B (c) subunit protein was immunoprecipitated. The same filters were stripped and successively probed with HRP-conjugated streptavidin (Strep, upper blot) and an antibody against the GluN1 (a), GluN2A (b) or GluN2B (c) subunit (lower blots). Bar graphs show summary data (mean ± SEM) of the normalized ratios between biotinylated and total GluN1, GluN2A or GluN2B subunit proteins detected. NMDAR phosphorylation induced by bath application of DHPG or N + G in neurons infected with PKD1shRNA is shown in (d and e). NMDAR phosphorylation in neurons infected with control shRNA (Ctl. shRNA) is shown in (f and g). The same filter shown in (d or e) was stripped and successively probed with anti-pS1416 (top blots), anti-pSer (middle blots) and GluN2A antibody (bottom blots). The anti-pSer (upper blots) and GluN2B antibody (lower blots) were respectively used to probe the filters shown in (e and g). Bar graphs show summary data (mean ± SEM) of the normalized ratios between phosphorylatd and total GluN2A (d and f) or GluN2B (e and g) protein. Open and filled bars in (d - g) show changes detected with anti-pSer and anti-pS1416 antibodies, respectively. #, ##: P < 0.05, 0.01 (independent t-test) in comparison with control

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