Figure 10. Expressing Pclo-Znf1-mCh rescues Pra1, EEA1 and Synaptophysin levels in Pclo^gt/gt neurons.

(A–C) Images of Pclo^wt/wt and Pclo^gt/gt neurons either expressing or not expressing Pclo-Znf1-mCh (arrows vs arrowheads) that were fixed and stained for Synaptophysin (A) and Pra1 (B) or EEA1 (C). (D) Quantification of (A). Pclo-Znf1-mCh slightly increases Synaptophysin levels in Pclo^wt/wt neurons (Pclo^wt/wt = 1 ± 0.01, n = 1497 synapses; Pclo^wt/wt + Pclo-Znf1-mCh = 1.16 ± 0.02, n = 710; four independent experiments). In Pclo^gt/gt boutons expressing Pclo-Znf1-mCh (arrows), Synaptophysin levels are increased compared to non-expressing boutons (arrowheads). (Pclo^gt/gt = 0.71 ± 0.01, n = 1203; Pclo^gt/gt + Pclo-Znf1-mCh = 1.12 ± 0.02, n = 590; four independent experiments). (E) Quantification of (B). Pclo-Znf1-mCh increases synaptic Pra1 levels in Pclo^wt/wt neurons (Pclo^wt/wt = 1 ± 0.04, n = 1136 synapses; Pclo^wt/wt + Pclo-Znf1-mCh = 1.31 ± 0.07, n = 543; 3–5 independent experiments). Pclo-Znf1-mCh rescues and further increases synaptic Pra1 levels in Pclo^gt/gt neurons (Pclo^gt/gt = 0.76 ± 0.05, n = 1064; Pclo^gt/gt + Pclo-Znf1-mCh = 1.80 ± 0.12, n = 445; 3–5 independent experiments). (F) Quantification of (C). Pclo-Znf1-mCh expression increases synaptic EEA1 levels in Pclo^wt/wt neurons (Pclo^wt/wt = 1 ± 0.03, n = 1652 synapses; Pclo^wt/wt + Pclo-Znf1-mCh = 1.22 ± 0.05, n = 709; 3–5 independent experiments). Pclo-Znf1-mCh rescues and further increases synaptic EEA1 levels in Pclo^gt/gt neurons (Pclo^gt/gt = 0.79 ± 0.03, n = 1633; Pclo^gt/gt + Pclo-Znf1-mCh = 1.24 ± 0.06, n = 464; 3–5 independent experiments). Scale bars represent 10 μm. Error bars in bar graph represent 95% confidence intervals. Numbers given represent mean ± SEM, ANOVA with Tukey multi comparison test. * denotes p<0.05, ** denotes p<0.01, *** denotes p<0.001 and **** denotes p<0.0001.