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. 2020 Oct 14;11:5171. doi: 10.1038/s41467-020-18956-x

Fig. 4. IgSF8 localizes to MF boutons and filopodia.

Fig. 4

a Cartoon of IgSF8 protein topology. b Western blot analysis of IgSF8 protein expression levels in mouse hippocampal homogenates at different developmental time points. Bar graph shows mean. Quantification from two independent experiments. c Subcellular fractionation using whole rat brains. H homogenate, CM crude membrane, Syn synaptosomes, Pre presynaptic fraction, PSD postsynaptic density fraction. d Western blot analysis of Igsf8 cKO mouse primary hippocampal cultures infected with a lentiviral vector harboring Cre recombinase or control vector. Bar graph shows mean ± SEM. Quantification from three independent experiments. e Confocal images of P28 Rbp4-Cre:Igsf8 cKO mouse hippocampal sections immunostained for IgSF8 and Synpr. Arrowheads indicate SL. Magnified insets of the SL in CA3 are shown on the right. f In utero electroporation setup to sparsely label DG granule cell neurons using membrane GFP (mGFP) and analyze the localization of HA-IgSF8 in MF synapses. g Stack of confocal images showing a mGFP-labeled MF bouton and respective filopodia (in blue in the merge). HA-IgSF8 is concentrated in the MF bouton, but also observed in defined regions along the filopodia, including their terminals (in green in the merge). More examples are shown in Supplementary Fig. 4e. Source data are provided as a Source data file. Scale bars in e 200 μm and in g 10 μm.