xph20 gfp  (Addgene inc)

Journal: bioRxiv

Article Title: Neurospheres from primary rodent brain cells to probe the 3D organization and function of synapses

doi: 10.64898/2026.03.19.712855

Figure Lengend Snippet: (A) Neurospheres were formed by mixing non-electroporated and electroporated cells before being plated in U-bottom ULA wells. (B) Maximum intensity projection image of a confocal stack of a DIV14 neurosphere in which neurons were separately electroporated with RFP (red) and Xph20-GFP (green). (C) Zoomed images on primary dendrites from neurons expressing intrabodies to PSD-95 (Xph20-GFP), or gephyrin (GPHN.FingR-GFP), revealing excitatory or inhibitory post-synapses, respectively. (D) Numbers of PSD-95 and gephyrin-positive puncta per electroporated neuron. Data represent the mean ± SEM of 14 and 10 neurospheres, respectively, and were compared by non-parametric Mann-Whitney test. Dots show individual neurospheres. (E) Maximum intensity projection of a confocal stack of a DIV14 neurosphere in which neurons were co-electroporated with GPHN.FingR-GFP (green) and Xph20-mRuby2 (magenta), allowing the detection of both excitatory and inhibitory post-synapses in the same cells. (F) Zoom on a dendritic segment corresponding to the rectangular area highlighted in (E). (G) Confocal image of a primary dendrite from a neuron expressing GFP-actin, further immunolabeled for GFP, showing numerous dendritic spines bulging out of the shaft.

Article Snippet: The intrabody to PSD-95, Xph20-GFP (Addgene #135,530 pCAG_Xph20-eGFP-CCR5TC), was a gift from M. Sainlos (IINS, University of Bordeaux) ( ).

Techniques: Expressing, MANN-WHITNEY, Immunolabeling

Journal: bioRxiv

Article Title: High-affinity detection of endogenously biotinylated neuroligin-1 at excitatory and inhibitory synapses using a tagged knock-in mouse strain

doi: 10.1101/2024.06.11.598408

Figure Lengend Snippet: (A) Dissociated neurons from bAP-NLGN1 hippocampal cultures were co-electroporated at DIV 0 with plasmids encoding BirA ER and two intrabodies (Xph20-mRuby2 and GPHN.FingR-GFP), which label respectively the excitatory and inhibitory post-synaptic proteins PSD-95 and gephyrin. The BirA ER plasmid has no reporter. (B) At DIV 14, neurons were live-labeled with SA-AF647 and visualized by epifluorescence microscopy. Representative image of a neuron showing bAP-NLGN1 labeled with SA-AF647 (magenta), PSD-95 positive synapses (cyan), and gephyrin positive synapses (green). An example of dendritic segment corresponding to the rectangle area is shown on the right. Insets showing zoomed synapses, with examples of SA-AF647 clusters overlapped with PSD- 95 (yellow arrows), or gephyrin (orange arrows) positive puncta. (C) Representative line scans across synaptic ROIs (insets) showing NLGN1 clusters either in an extrasynaptic location (ExtraSyn), overlapping with PSD-95 (PSD-95+), overlapping with gephyrin (GPHN+), or with both PSD-95 and gephyrin (PSD-95+/GPHN+). (D) Representation of bAP-NLGN1 cluster distribution throughout the dendrite. (E, F) Bar plots representing the enrichment of bAP-NLGN1 at PSD-95 or gephyrin positive puncta, and the fraction of these puncta overlapping with bAP-NLGN1, respectively. Dots in the bars represent individual cells, with n = 39 cells for each condition, from 2 independent experiments. Data represent mean ± SEM and were compared by a Mann-Whitney test (*P < 0.05, ***P < 0.001). (G) Violin plots showing the surface area of individual bAP-NLGN1 clusters as a function of their localization. Quantification was performed either on all NLGN1 clusters present throughout the dendritic segment (All), on clusters overlapping with PSD-95 or gephyrin (PSD-95+/GPHN+), or on clusters localized at extrasynaptic sites (ExtraSyn) (n = 436-2224 clusters in 39 cells, from 2 independent experiments). Data were compared by a Kruskal–Wallis test followed by Dunn’s multiple comparison test (****P < 0.0001).

Article Snippet: The intrabodies to PSD-95 , Xph20-GFP and Xph20-mRuby2 (Addgene #135,530 pCAG_Xph20-eGFP-CCR5TC; #135,531 pCAG_Xph20-mRuby2- CCR5TC), were gifts from M. Sainlos (IINS, University of Bordeaux).

Techniques: Plasmid Preparation, Labeling, Epifluorescence Microscopy, MANN-WHITNEY, Comparison