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

. 2022 Dec 10;13:7656. doi: 10.1038/s41467-022-35417-9

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

© The Author(s) 2022

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. 5 — A Schematic illustrating the possibility that the GluRIIA C-tail stabilizes active pCaMKII at postsynaptic compartments. B Amino acid alignment of the mouse CaMKII autoinhibitory domain and homologous region in Drosophila, with the interaction sequences encoded in the C-tails of mouse GluN2B, Drosophila EAG, and the putative CaMKII interaction domain in the Drosophila GluRIIA C-tail. The black arrow indicates the Thr residue that is phosphorylated in active pCaMKII. The red Arg residue indicates the necessary R-X-X-S motif; dark gray indicates strongly conserved while light gray indicates weakly conserved residues. C Diagram of the GluRIIA C-tail amino acid sequence and three mutant truncation alleles of the C-tail induced by CRISPR mutagenesis. The regions of the two guide RNAs used to generate these alleles are shown, as well as the antigenic domains of two GluRIIA-specific antibodies. D Representative images of NMJ boutons immunostained with anti-GluRIIA, -GluRIIA^tail, -GluRIIC, -pCaMKII, -CaMKII, and -DLG antibodies in the indicated genotypes: GluRIIA^ΔC20 (w;GluRIIA^ΔC20), GluRIIA^ΔC9 (w;GluRIIA^ΔC9), GluRIIA^QRΔC19, and (w;GluRIIA^QRΔC19). Experiments were repeated three times independently with similar results. Note that the loss of the terminal six amino acids of the GluRIIA C-tail is sufficient to completely lose pCaMKII signals. E Quantification of mean fluorescence intensities of the indicated antibody signal normalized to wild-type values (wild type: n = 12; GluRIIA^−/−, n = 12, p < 0.0001 for GluRIIA, p < 0.0001 for GluRIIA^tail, p = 0.9912 for GluRIIC, p < 0.0001 for pCaMKII, p = 0.9958 for CaMKII, p = 0.0022 for DLG; GluRIIA^ΔC20: n = 12, p = 0.1744 for GluRIIA, p < 0.0001 for GluRIIA^tail, p = 0.9912 for GluRIIC, p < 0.0001 for pCaMKII, p = 0.9958 for CaMKII, p = 0.0022 for DLG; GluRIIA^ΔC6: n = 12, p = 0.6019 for GluRIIA, p < 0.0001 for GluRIIA^tail, p = 0.9912 for GluRIIC, p < 0.0001 for pCaMKII, p = 0.9958 for CaMKII, p = 0.0022 for DLG; GluRIIA^QRΔC19: n = 12, p = 0.9967 for GluRIIA, p < 0.0001 for GluRIIA^tail, p = 0.9912 for GluRIIC, p < 0.0001 for pCaMKII, p = 0.9958 for CaMKII, p = 0.0022 for DLG). Error bars indicate ±SEM. *p < 0.05, ****p < 0.0001; ns, not significant. n values indicate biologically independent NMJs. Source data are provided as a Source Data file.