High-efficiency localization of Na+–K+ ATPases on the cytoplasmic side by direct stochastic optical reconstruction microscopy

Abstract

We describe a concise and effective strategy towards precisely mapping Na⁺–K⁺ ATPases on the cytoplasmic side of cell membranes by direct stochastic optical reconstruction microscopy (dSTORM). We found that most Na⁺–K⁺ ATPases are localized in different sizes of clusters on human red blood cell (hRBC) membranes, revealed by Ripley's K-function analysis. Further evidence that cholesterol depletion causes the dispersion of Na⁺–K⁺ ATPase clusters indicates that such clusters could be localized in cholesterol-enriched domains. Our results suggest that Na⁺–K⁺ ATPases might aggregate within the lipid rafts to fulfill their functions.