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. 2020 May 19;295(27):9244–9262. doi: 10.1074/jbc.RA120.013077

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© 2020 Kim et al.

Published under exclusive license by The American Society for Biochemistry and Molecular Biology, Inc.

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Figure 8. — Interactions of Clstn3 via its cadherin domains with Nrxns are involved in Clstn3-mediated regulation of specific excitatory Schaffer-collateral projections in the CA1 hippocampal region. A, experimental protocols for quantitative immunohistochemistry experiments presented in B–E. Quantitative immunohistochemical analyses were performed 2 weeks after stereotactic injection of the indicated AAVs into the CA1 hippocampal regions of ∼9-week–old Nestin-Clstn3 mice. B and D, representative images of hippocampal CA1 SO, SR, and SLM regions 2 weeks after stereotactic injections of the indicated AAVs into Clstn3^fl/fl (Ctrl) or Nestin-Clstn3 mice, followed by immunostaining for the excitatory synapse marker VGLUT1 (B) or PSD-95 (D). Scale bar: 20 μm (applies to all images). C and E, quantification of the integrated intensity of VGLUT1-positive (C) or PSD-95–positive (E) synaptic puncta. Data are mean ± S.E. (*, p < 0.05; **, p < 0.01; ***, p < 0.001; ANOVA with Tukey's test; n = 5 mice each after averaging data from 4 to 6 sections/mouse). p values for VGLUT1 in CA1 SO: Ctrl versus Nestin-Clstn3, p = 0.0075; Ctrl versus Clstn3 Full (rescue), p > 0.9999; Ctrl versus Clstn3 ΔCad (rescue), p = 0.0166; Ctrl versus Clstn3 Cad (rescue), p > 0.9999; Nestin-Clstn3 versus Clstn3 full (rescue), p = 0.0011. p values for VGLUT1 in CA1 SR: Ctrl versus Nestin-Clstn3, p = 0.0021; Ctrl versus Clstn3 full (rescue), p = 0.7147; Ctrl versus Clstn3 ΔCad (rescue), p = 0.0028; Ctrl versus Clstn3 Cad (rescue), p = 0.9921; Nestin-Clstn3 versus Clstn3 Full (rescue), p = 0.0359. p values for VGLUT1 in CA1 SLM: Ctrl versus Nestin-Clstn3, p > 0.9999; Ctrl versus Clstn3 Full (rescue), p > 0.9999; Ctrl versus Clstn3 ΔCad (rescue), p > 0.9999; Ctrl versus Clstn3 Cad (rescue), p > 0.9999. p values for PSD-95 in CA1 SO: Ctrl versus Nestin-Clstn3, p = 0.0266; Ctrl versus Clstn3 full (rescue), p = 0.4300; Nestin-Clstn3 versus Clstn3 Full (rescue), p = 0.0009. p values for PSD-95 in CA1 SR: Ctrl versus Nestin-Clstn3, p = 0.0002; Ctrl versus Clstn3 Full (rescue), p = 0.8762; Nestin-Clstn3 versus Clstn3 Full (rescue), p = 0.0010. p values for PSD-95 in CA1 SLM: Ctrl versus Nestin-Clstn3, p = 0.9397; Ctrl versus Clstn3 Full (rescue), p = 0.3147; Nestin-Clstn3 versus Clstn3 Full (rescue), p = 0.4975.