Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 2021 May 24;12:3057. doi: 10.1038/s41467-021-23116-w

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

© The Author(s) 2021

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

PMC Copyright notice

Fig. 4 — a Schematic for simultaneous knockout of Liprin-α2 and -α3. Mice in which Liprin-α2 can be removed by cre recombination (Liprin-α2^f/f) were generated and crossed to previously published constitutive Liprin-α3 knockout mice (Liprin-α3^−/−, generated by CRISPR/Cas9-mediated genome editing, deleted sequence is represented with dashes)¹⁹. Cultured hippocampal neurons of Liprin-α2^f/f/Liprin-α3^−/− mice infected with lentivirus expressing cre recombinase (KO^L23) were compared to neurons of Liprin-α2^f/f x Liprin-α3^+/− mice infected with lentiviruses that express an inactive cre recombinase (control^L23). b, c Example confocal images (b) and quantification (c) of neurons immunostained for Liprin-α2, Liprin-α3, RIM, Munc13-1, RIM-BP2, Bassoon, Ca_V2.1, PSD-95, Gephyrin, or GluA1 along with Synaptophysin (for Munc13-1, RIM-BP2, and Gephyrin) or Synapsin (all others) as vesicle marker, or Synapsin-1 and MAP2. Quantification in c was performed in regions of interest (ROIs) defined by the synaptic vesicle marker and normalized to the average control^L23 levels per culture, N = 30 images/3 independent cultures per genotype per protein of interest, except for Munc13-1 (N = 15/3), RIM-BP2 (N = 14/3), and Gephyrin (N = 14/3), p values: Liprin-α2, 2e-16 (***); Liprin-α3, 2e-16 (***); RIM, 3e-11 (***); Munc13-1, 3e-05 (***); RIM-BP2, 0.54; Bassoon, 0.07; PSD-95, 0.30; Gephyrin, 0.10; GluA1, 0.53; Ca_V2.1, 5e-10 (***); Synapsin, 7e-06 (***). d–h Example electron micrographs of synapses (d) and quantification (per section) of the number of synaptic vesicles (e), bouton size (f), PSD length (g), and number of docked vesicles (h) of neurons fixed by high-pressure freezing followed by freeze substitution, control^L23: N = 111 synapses/2 independent cultures, KO^L23: N = 124/2, p values: e, 0.02 (*); f, 0.51; g, 0.95; h, 0.02 (*). All data were mean ± SEM. Significance was assessed using Mann–Whitney U tests, except for PSD-95 and Gephyrin in c, for which t-tests were used. All tests were two-sided. For synaptic localization of Liprin-α1 to -α4, see Supplementary Fig. 7, for generation of Liprin-α2^f/f mice and analysis of Synaptophysin levels of the experiments shown in c, see Supplementary Fig. 8.