Integrating GlycoSHIELD Modeling and DNA-PAINT SMLM to Map the Glycosylation-Dependent Distribution of the Na,K-ATPase

This dataset accompanies the study "Integrating GlycoSHIELD Modeling and DNA-PAINT SMLM to Map the Glycosylation-Dependent Distribution of the Na,K-ATPase" (Stojcic et al., Journal of Membrane Biology, 2026; https://doi.org/10.1007/s00232-026-00396-1). The dataset contains raw single-molecule localization microscopy (SMLM) data and plasmid sequence data generated to investigate how N-glycosylation of the Na,K-ATPase β1 subunit governs nanoscale pump organization at the plasma membrane. SMLM data were acquired using DNA-PAINT under TIRF (Total Internal Reflection Fluorescence) illumination on a Carl Zeiss Elyra PS.1 microscope. A498 cells expressing either wild-type (WT) or glycosylation-deficient (3NQ) Na,K-ATPase β1-GFP constructs were imaged across three biological replicates (6–7 cells per condition). Localization coordinates were used for Ripley's H-function and DBSCAN (Density-Based Spatial Clustering of Applications with Noise) cluster analysis. Plasmid sequence data document the site-directed mutagenesis of human ATP1B1-GFP, in which asparagine residues at positions 158, 193, and 265 were substituted with glutamine to generate the 3NQ glycosylation-deficient mutant. Analysis code is available at github.com/bruno-stojcic/glyco-PAINT-analysis.