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. 2013 Jul 17;33(29):12122–12135. doi: 10.1523/JNEUROSCI.0131-13.2013

Figure 4.

Figure 4.

Phospho-null and phosphomimic mutations of T19 alter PSD-95 stability in dendritic spines. A–C, Sample images of dendritic spines from CA1 pyramidal neurons expressing DsRed and (A) WT-PSD-95, (B) T19A-PSD-95, or (C) T19D-PSD-95 fused to photoactivatable GFP. Vertical dashed lines indicate initial activation laser and postexperimental reactivation with the 405 nm laser. D, Graph shows time course of PAGFP fluorescence (mean ± SEM) from individual spines of neurons expressing PSD-95-PAGFP constructs as indicated. Statistical significance for the T19A and T19D mutants was determined by two-way ANOVA with a Bonferroni post hoc test with comparisons to wild-type PSD-95-PAGFP at each time point (*p < 0.05, **p < 0.01, ***p < 0.001). Wild-type PSD-95-PAGFP, n = 18 spines; T19A-PSD-95-PAGFP, n = 13 spines; T19D-PSD-95-PAGFP, n = 14 spines. E, Time courses of PAGFP fluorescence for the PSD-95-T19/S295-PAGFP double phosphomutants. Data for wild-type PSD-95-PAGFP is replotted from D for reference. F, PAGFP intensities at the 55 min time point for the T19, S295, and T19/S295 mutants of PSD-95, compared with wild-type PSD-95 (gray bar). Statistical analysis was performed by one-way ANOVA, followed by a Bonferroni analysis comparing all treatments with wt-PSD-95-PAGFP (n = 18, 13, 14, 11, 12, 14, 11, 15, and 13 spines from left to right; *p < 0.05, **p < 0.01, ***p < 0.001).