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. 2020 Oct 28;40(44):8438–8462. doi: 10.1523/JNEUROSCI.1091-20.2020

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Figure 2. — PTPσ is required for timed presynaptic differentiation. A, B, Representative images of heterologous synapse-formation assays. Cultured hippocampal neurons were infected with the indicated lentiviruses at DIV4, and cocultured with mVenus-fused Nlgn1 (A) or Slitrk6-expressing (B) HEK293T cells at DIV12 for 6, 12, or 24 h. Synaptogenic activities were analyzed by double-immunostaining for HA/EGFP (blue) and VGLUT1 (red). Scale bar, 10 μm. For additional results, see Extended Data Figure 2-1. C, D, Synapse-formation activities in A and B were quantified by measuring the ratio of VGLUT1 staining intensity (red) to HA/EGFP intensity (blue). Data are mean ± SEM. *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001; ANOVA with a nonparametric Kruskal–Wallis test. n indicates the number of cells as follows: sh-Control/Nlgn1 (6 h), n = 14; sh-PTPσ/Nlgn1 (6 h), n = 13; sh-PTPδ/Nlgn1 (6 h), n = 12; sh-Nrxns/Nlgn1 (6 h), n = 14; sh-Control/Slitrk6 (6 h), n = 10; sh-PTPσ/Slitrk6 (6 h), n = 11; sh-PTPδ/Slitrk6 (6 h), n = 13; and sh-Nrxns/Slitrk6 (6 h), n = 11; sh-Control/Nlgn1 (12 h), n = 13; sh-PTPσ/Nlgn1 (12 h), n = 14; sh-PTPδ/Nlgn1 (12 h), n = 12; sh-Nrxns/Nlgn1 (12 h), n = 13; sh-Control/Slitrk6 (12 h), n = 12; sh-PTPσ/Slitrk6 (12 h), n = 11; sh-PTPδ/Slitrk6 (12 h), n = 11; sh-Nrxns/Slitrk6 (12 h), n = 11; sh-Control/Nlgn1 (24 h), n = 14; sh-PTPσ/Nlgn1 (24 h), n = 13; sh-PTPδ/Nlgn1 (24 h), n = 14; sh-Nrxns/Nlgn1 (24 h), n = 14; sh-Control/Slitrk6 (24 h), n = 15; sh-PTPσ/Slitrk6 (24 h), n = 12; sh-PTPδ/Slitrk6 (24 h), n = 13; and sh-Nrxns/Slitrk6 (24 h), n = 12; sh-Control/Control (24 h), n = 10; sh-PTPσ/Control (24 h), n = 10; sh-PTPδ/Control (24 h), n = 9; sh-Nrxns/Control (24 h), n = 11. p values for each comparison are as follows: sh-Control versus sh-PTPσ/Nlgn1 (6 h), p > 0.9999; sh-Control versus sh-PTPδ/Nlgn1 (6 h), p > 0.9999; sh-Control versus sh-Nrxns/Nlgn1 (6 h), p > 0.9999; sh-Control versus sh-PTPσ/Slitrk6 (6 h), p = 0.0575; sh-Control versus sh-PTPδ/Slitrk6 (6 h), p > 0.9999; and sh-Control versus sh-Nrxns/Slitrk6 (6 h), p > 0.9999; sh-Control versus sh-PTPσ/Nlgn1 (12 h), p = 0.0002; sh-Control versus sh-PTPδ/Nlgn1 (12 h), p = 0.2086; sh-Control versus sh-Nrxns/Nlgn1 (12 h), p < 0.0001; sh-Control versus sh-PTPσ/Slitrk6 (12 h), p > 0.9999; sh-Control versus sh-PTPδ/Slitrk6 (12 h), p > 0.9999; sh-Control versus sh-Nrxns/Slitrk6 (12 h), p > 0.9999; sh-Control versus sh-PTPσ/Nlgn1 (24 h), p > 0.9999; sh-Control versus sh-PTPδ/Nlgn1 (24 h), p = 0.5651; sh-Control versus sh-Nrxns/Nlgn1 (24 h), p = 0.0071; sh-Control versus sh-PTPσ/Slitrk6 (24 h), p = 0.0215; sh-Control versus sh-PTPδ/Slitrk6 (24 h), p = 0.8652; and sh-Control versus sh-Nrxns/Slitrk6 (24 h), p > 0.9999; sh-Control versus sh-PTPσ/Control (24 h), p = 0.6487; sh-Control versus sh-PTPδ/Control (24 h), p = 0.6644; sh-Control versus sh-Nrxns/Control (24 h), p > 0.9999.