Fig. 6: Validation of R-mAbs and i-mAbs for labeling synaptic AMPARs.
From: Genetically encoded intrabody probes for labeling and manipulating AMPA-type glutamate receptors
a STED images show primary hippocampal dendrites from GFP-expressing neurons (red) labeled with A1 IgG antibody (left), A1 scFv R-mAb (middle) or A1 scFv i-mAb (right) (cyan) along with immunostaining for PSD95 (yellow). b Higher magnification images show individual dendritic spines labeled with anti-PSD95 (yellow), anti-Homer1 (red) or conventional anti-GluA1, A1 scFv-Halo R-mAb, or A1 scFv-Halo i-mAb (blue) and 3D reconstructions of the same synapses. c Fraction of A1 intensity inside PSD95 nanodomain is plotted (anti-A1 n = 147 synapses from 6 cells from 2 cultures, A1 R-mAb n = 163 from 5 cells from 1 culture, A1 i-mAb n = 121 from 6 cells from 2 cultures, Dunnett’s multiple comparisons test, anti-A1 vs. A1 R-mAb n.s. p = 0.7873, anti-A1 vs. A1 i-mAb n.s. p = 0.0539). d STED images of primary hippocampal dendrites expressing GFP (red) and labeled with anti-PSD95 (yellow) and either A2 IgG antibody (left), A2 Fab-Halo R-mAb (middle) or A2 scFv-Halo R-mAb (right) (cyan). e Higher magnification images show individual dendritic spines labeled with anti-PSD95 (yellow), anti-Homer1 (red) or conventional anti-GluA2, A2 Fab-Halo R-mAb, or A2 scFv-Halo R-mAb (blue) and 3D reconstructions of the same synapses. f Plot of fraction of A2 intensity inside PSD95 nanodomain (anti-A2 n = 88 synapses from 6 cells from 2 cultures, A2 Fab-Halo R-mAb n = 162 from 8 cells from 2 cultures, A2 scFv-Halo R-mAb n = 131 from 6 cells from 1 culture, Dunnett’s multiple comparisons test, anti-A2 vs. A2 Fab-Halo R-mAb n.s. p = 0.3927, anti-A2 vs. A2 scFv-Halo R-mAb n.s. p = 0.3756). Data are presented as mean values +/- SEM. n.s. not significant. Source data are provided as a Source Data file.
