Fig. 4

Local knockdown of IgSF9b in the adult CeM ameliorates anxiety-related behaviors in Nlgn2 KO mice. a Immunoblot for IgSF9b in HEK cell lysates transfected with a Myc-IgSF9b construct and IgSF9b shRNA or control shRNA¹⁴. Numbers next to the protein ladder represent molecular weight in kDa. b Representative images and quantification of perisomatic IgSF9b puncta in the CeM following injection of control shRNA or IgSF9b shRNA. n = 3 mice. Scale bar, 20 μm. c Schematic diagram showing experimental design of IgSF9b shRNA experiment. Mice were tested in the OF 24 h before (pre) and 6 weeks after (post) injection. d Representative image of GFP-positive neurons in viral injection site. e Four experimental groups were generated: WT + control shRNA (white), WT + IgSF9b shRNA (white and blue striped), Nlgn2 KO + control shRNA (red), and Nlgn2 KO + IgSF9b shRNA (red and blue striped). f Time in center post-injection expressed as % pre-injection. g Normalized distance in center post-injection expressed as % pre-injection. h Representative tracks of OF exploration of Nlgn2 KO mice pre- and post-injection. Statistically significant ANOVA comparisons are marked in gray at the top of the panels and are listed in Table 1. For all other ANOVA comparisons F < 1. Post-hoc analysis: *p < 0.05 relative to WT, **p < 0.01 relative to WT. Error bars represent SEM, n = 6–9 mice per group