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. 2016 Oct 20;9:104. doi: 10.3389/fnmol.2016.00104

FIGURE 7.

FIGURE 7

Impaired synaptogenic activity of a subset of Slitrk mutants in cultured hippocampal neurons. (A-C) Representative images of the heterologous synapse-formation activities of WT Slitrk and the indicated point mutants in cultured hippocampal neurons. Neurons were cocultured with HEK293T cells transfected with WT or the indicated mutant forms of HA-Slitrk1 (A), HA-Slitrk2 (B), or HA-Slitrk4 (C) at DIV10. Neurons at DIV12 were then immunostained with antibodies against EGFP or HA (blue) and synapsin (red). Scale bar, 10 μm (applies to all images). (D) Synapse-formation activity in panels (A–C) was quantified by measuring the ratio of synapsin staining intensity (red) to HA/EGFP intensity (blue). All data are shown as means ± SEMs (∗p < 0.05; 2∗p < 0.01; 3∗p < 0.001; ANOVA with post hoc Tukey’s test; p-value of WT Slitrk1 vs. Slitrk1[N400I] = 0.00176; p-value of WT Slitrk1 vs. Slitrk1[T418S] = 0.0497; p-value of WT Slitrk1 vs. Slitrk1[R584K] = 0.959; p-value of WT Slitrk1 vs. Slitrk1[S593G] = 0.434; p-value of WT Slitrk2 vs. Slitrk2[R32L] = 0.818; p-value of WT Slitrk2 vs. Slitrk2[V89M] = 0.957; p-value of WT Slitrk2 vs. Slitrk2[S549F] = 0.956; p-value of WT Slitrk2 vs. Slitrk2[S601P] = 0.984; p-value of WT Slitrk2 vs. Slitrk2[L626F] = 0.987; p-value of WT Slitrk4 vs. Slitrk4[V206I] = 0.00062; and p-value of WT Slitrk4 vs. Slitrk4[I578V] = 1.5112E-7). ‘n’ denotes the number of HEK293T cells as follows: Control, n = 14; WT Slitrk1, n = 14; Slitrk1[N400I], n = 12; Slitrk1[T418S], n = 14; Slitrk1[R584K], n = 14; Slitrk1[S593G], n = 21; WT Slitrk2, n = 12; Slitrk2[R32L], n = 10; Slitrk2[V89M], n = 10; Slitrk2[S549F], n = 15; Slitrk2[S601P], n = 15; Slitrk2[L626F], n = 15; WT Slitrk4, n = 17; Slitrk4[V206I], n = 13; and Slitrk4[I578V], n = 15.