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

FIGURE 8.

FIGURE 8

Impaired regulation of synapse density by a subset of Slitrk mutants in cultured hippocampal neurons. (A) Cultured hippocampal neurons were transfected with a lentiviral vector expressing sh-Control, sh-Slitrk1, sh-Slitrk2 or sh-Slitrk4, or coexpressing the indicated sh-Slitrk vector and corresponding shRNA-resistant Slitrk vectors at DIV8 and analyzed at DIV14 by double-immunofluorescence staining with antibodies to EGFP (green) and synapsin (red). Scale bar, 5 μm (applies to all images). (B,C) Summary data showing the effects of Slitrk molecular replacement in neurons on synapsin puncta density (B) and synapsin puncta size (C). More than three dendrites per transfected neuron were analyzed and group-averaged. All data shown are means ± SEMs (∗p < 0.05; 2∗p < 0.01; 3∗p < 0.001; ANOVA with post hoc Tukey’s test; p-value of sh-Control vs. sh-Slitrk1 = 0.029; p-value of sh-Control vs. sh-Slitrk1+WT Slitrk1 = 0.99; p-value of sh-Control vs. sh-Slitrk1+Slitrk1[N400I] = 0.004; p-value of sh-Control vs. sh-Slitrk1+Slitrk1[T418S] = 0.049; p-value of sh-Control vs. sh-Slitrk2 = 0.039; p-value of sh-Control vs. sh-Slitrk2+WT Slitrk2 = 0.99; p-value of sh-Control vs. sh-Slitrk2+Slitrk2[V89M] = 0.0093; p-value of sh-Control vs. sh-Slitrk4 = 0.00075; p-value of sh-Control vs. sh-Slitrk4+WT Slitrk4 = 0.877; p-value of sh-Control vs. sh-Slitrk4+Slitrk4[V206I] = 0.003; and p-value of sh-Control vs. sh-Slitrk4+Slitrk4[I578V] = 0.008). ‘n’ denotes the number of neurons as follows: sh-Control, n = 16, sh-Slitrk1, n = 14, sh-Slitrk1+WT Slitrk1, n = 14; sh-Slitrk1+Slitrk1[N400I], n = 14; sh-Slitrk1+Slitrk1[T418S], n = 14; sh-Slitrk2, n = 13; sh-Slitrk2+WT Slitrk2, n = 13; sh-Slitrk2+Slitrk2[V89M], n = 13; sh-Slitrk4, n = 17; sh-Slitrk4+WT Slitrk4, n = 15; sh-Slitrk4+Slitrk4[V206I], n = 15; and sh-Slitrk4+Slitrk4[I578V], n = 15.