Data and code from: Tetraspanin CD82 shapes EGFR signaling outcomes through nanoscale receptor organization
Data files
Mar 03, 2026 version files 11.17 MB
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20251020_Fig1B-C_HeLa_dSTORM_ROIs.xlsx
14.42 KB
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20251020_Fig1G-I_A431_dSTORM_EGFR-Clusters.xlsx
63.01 KB
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20251020_Fig1G-I_HEK293_dSTORM_EGFR-Clusters.xlsx
97.37 KB
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20251020_Fig2D_A431_BaGoL_CD82-Clusters.xlsx
191.29 KB
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20251110_Fig3_A431_BaGoL_EGFR-Clusters.xlsx
10.79 MB
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20251126_Final-Analyses-and-Scripts.zip
10.82 KB
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README.md
3.70 KB
Abstract
Tetraspanins are integral membrane proteins that play a crucial role in organizing and regulating cellular signaling by serving as scaffolds that compartmentalize receptors and other signaling molecules within membrane microdomains. Here, we report how the tetraspanin CD82 modulates the molecular organization and signaling of the Epidermal Growth Factor Receptor (EGFR), a key molecule involved in cellular proliferation, differentiation, and survival. Combining multi-color super-resolution microscopy with advanced image reconstruction and analysis techniques, we demonstrate that CD82 selectively associates with EGFR, promotes receptor oligomerization, and acts as a regulator of ligand-independent receptor phosphorylation in a palmitoylation-dependent manner. Additionally, CD82 promotes a more compact molecular organization of EGFR, which correlates with altered endocytosis and downstream signaling outcomes. These findings underscore the importance of tetraspanins in the spatial and functional regulation of cell surface receptors, with implications for controlling aberrant signaling in disease and positions CD82 as a potential target for developing therapeutic strategies aimed at modulating EGFR signaling by influencing receptor organization.
Dataset DOI: 10.5061/dryad.3n5tb2rz2
Description of the data and file structure
Data used for quantifying statistically significant differences in the molecular organization of EGFR are depicted in Figures 1 - 3. Statistical analyses were accomplished using generalized linear mixed effects models to estimate and compare differences between treatments/cell lines while accounting for the nested nature of the SR imaging datasets (with clustering metrics potentially being gathered from several ROIs within a cell, and potentially multiple cells being imaged per day). All underlying data, as well as code used for analysis of the data, can be found here.
Files and variables
File: 20251020_Fig1B-C_HeLa_dSTORM_ROIs.xlsx
Description: Values of Hopkins Statistics and relative number of clusters for EGFR in HeLa cells, as depicted in Figure 1B-C. Includes data on the corresponding imaging dates, treatments, cells, and ROI number for each data point.
Variables
- TotalLocalizations, # of Clusters, Relative # of Clusters, Hopkins Statistic, Imaging Date, Cell line, Treatment, Cell #, ROI #
File: 20251020_Fig1G-I_A431_dSTORM_EGFR-Clusters.xlsx
Description: Values of Hopkins Statistics and relative number of clusters for EGFR in A431 cells, as depicted in Figure 1G-I. Includes data on the corresponding imaging dates, treatments, cells, and ROI number for each data point.
Variables
- TotalLocalizations, # of Clusters, Relative # of Clusters, Hopkins Statistic, Imaging Date, Cell line, Treatment, Cell #, ROI #
File: 20251020_Fig1G-I_HEK293_dSTORM_EGFR-Clusters.xlsx
Description: Values of Hopkins Statistics and relative number of clusters for EGFR in HEK293 cells, as depicted in Figure 1G-I. Includes data on the corresponding imaging dates, treatments, cells, and ROI number for each data point.
Variables
- TotalLocalizations, # of Clusters, Relative # of Clusters, Hopkins Statistic, Imaging Date, Cell line, Treatment, Cell #, ROI #
File: 20251020_Fig2D_A431_BaGoL_CD82-Clusters.xlsx
Description: Values of Percent of points clustered, ROI-averaged cluster sizes, ROI-averaged points per clusters, as well as sizes and points per cluster for all clusters, for BaGoL-processed imaging data of CD82 in A431 cells, as depicted in Figure 2C-D. Includes data on the corresponding imaging dates, cells, and ROI number for each data point.
Variables
- Num Clustered Locs, Total Localizations, Percent Clustered, ROI Avg Cluster Size (Equiv Radii), ROI Avg Pts per Cluster, PtsPerCluster, Equiv Radii, Imaging Date, Cell line, Cell #, ROI #
File: 20251110_Fig3_A431_BaGoL_EGFR-Clusters.xlsx
Description: Values of Percent of points clustered, # of clusters, ROI-averaged cluster diameters, as well as distributions of EGFR clusters, and percentages of EGFR found in dimeric or oligomeric clusters, for BaGoL-processed imaging data of EGFR in A431 cells, as depicted in Figure 3B-G. Includes data on the corresponding imaging dates, cells, and ROI number for each data point.
Variables
- Total Localizations, Num Clustered Locs, Percent Clustered, Number of Clusters, Relative # Clusters, MedianClusterDiameter, PctEGFRinDimers, PctEGFRinOligomers, ClustFract_2pt, ClustFract_3pt, ClustFract_4pt, ClustFract_5pt+, ClusterDiameter, PtsPerClust, Imaging Date, Cell line, Treatment, Cell #, ROI #
File: 20251126_Final-Analyses-and-Scripts.zip
Description: Annotated SAS code used for analyses of data described above.