Journal: Channels
Article Title: Calmodulin binds to the N-terminal domain of the cardiac sodium channel Na v 1.5
doi: 10.1080/19336950.2020.1805999
Figure Lengend Snippet: WT and variant channel-ER colocalization is similar in homo- and heterozygous conditions. (a) Airyscan microscopy images of RNCs transiently co-transfected with WT or variant flag-Na v 1.5 (green) and the ER marker calreticulin-DsRed2 (red), representing homologous conditions. Nuclei are stained with DAPI (blue). Right column: merged images. (b) Airyscan microscopy images of RNCs transiently co-transfected with GFP-Na v 1.5 WT (green), flag-Na v 1.5 WT or variants (not shown), and calreticulin-DsRed2 (red). Right column: merged images; nuclei are stained with DAPI (blue). (c) Quantification of Na v 1.5-ER colocalization in homozygous conditions. Colocalization is defined as percentage of Na v 1.5 signal area colocalizing with ER signal area within predefined ROI. (d) Quantification of colocalization of WT-Na v 1.5 with ER in the presence of variants, representing heterozygous conditions. Cell areas in which Na v 1.5 and ER colocalize are: WT, 57.4 ± 9.3%; Y87C, 54.0 ± 12.6%; R104W, 75.1 ± 9.5%; and R121W, 47.9 ± 10.5%. Scale bar, 10 µm. Data are presented as mean ± SEM. Groups were not significantly different.
Article Snippet: We used ZEN 2.1 software for processing raw Airyscan data, and the IMARIS coloc tool (IMARIS 9.3.1 software, Bitplane, Zürich, Switzerland) to quantify the colocalization between signals of two channels within the predefined ROI (region of interest, defined as the cell area).
Techniques: Variant Assay, Microscopy, Transfection, Marker, Staining