Fig. 6

Postsynaptic changes in netrin-G KO mice. a and b Layer selective distribution of NGL1 in the hippocampal CA1 of netrin-G1 KO and WT mice. a Western blot images for NGL1 and actin in the SLM and SR samples. b Relative intensities of NGL1 normalized to actin (sample = animal numbers are indicated in the columns) indicate diffusion of NGL1 across layers of netrin-G1 KO mice. c and d Layer-specific distribution of NGL2 in the hippocampal CA1 of netrin-G2 KO and WT mice. C, Western blot images for NGL2 and actin in the SLM and SR samples. D, Relative intensities of NGL2 normalized to actin (sample = animal numbers are indicated in the columns) indicate diffusion of NGL2 across layers of netrin-G2 KO mice. e-j Confocal microscopy images obtained after dual immunohistochemistry for PSD-95 (green) and NGL-2 (red) in the cortex layer 4 of WT (e-g) and netrin-G2 KO (h-j) mice revealed that NGL-2 was co-localized with a postsynaptic marker in both WT and netrin-G2 KO mice (scale bar: 2 μm). k and l Western blot immunoassay for NGLs in synaptoneurosome fractions of WT, netrin-G1 KO, and netrin-G2 KO mice revealed no significant differences among genotypes (sample = animal numbers are indicated in the columns). m and n Western blot immunoassay for NGLs in CSC and SPM fractions of WT, netrin-G1 KO, and netrin-G2 KO mice. N: The relative intensities of NGLs normalized to β-tubulin were compared in SPM fractions (sample = animal numbers are indicated in the columns; 1-way ANOVA, * < 0.05 post-hoc Scheffe’s test). Although the sub-synaptic localization of NGL-1 was not altered in netrin-G1 KO mice, the sub-synaptic localization of NGL-2 was altered in netrin-G2 KO mice. The decrease in NGL-2 in the SPM fraction was associated with a marked increase in NGL-2 in the CSC fraction in netrin-G2 KO mice. Data are presented as mean ± SEM