The PDF file includes:

• Fig. S1. The multistep paradigm of cell migration highlighting the main interactions that take place between the blood stem cell in flow and the endothelial cells lining the blood vessels of the bone marrow.
• Fig. S2. Surface density of E-selectin on the protein A–deposited microfluidic chamber determined by immunofluorescence imaging.
• Fig. S3. Flow cytometric analysis to determine the binding specificity of ligand-specific antibodies to CD44 on KG1a cells.
• Fig. S4. Localization precisions of the SR localization microscopy experiments of CD44 on KG1a cells.
• Fig. S5. Examples of the reconstructed SR images of CD44 on KG1a cells.
• Fig. S6. SR images of CD44 on KG1a cells.
• Fig. S7. Cluster analysis of the nanoscale architecture of lipid rafts on KG1a cells.
• Fig. S8. Examples of the reconstructed SR images of CD44 on MβCD-treated KG1a cells.
• Fig. S9. Cluster analysis of the nanoscale architecture of CD44 on KG1a cells.
• Fig. S10. Expression of CD44 on untreated and MβCD-treated KG1a cells was determined by flow cytometry.
• Fig. S11. Depth of the field in the SR localization microscopy imaging experiments with HILO configuration.
• Legends for movies S1 to S8

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Other Supplementary Material for this manuscript includes the following:

• Movie S1 (.mp4 format). Time-lapse transmitted light microscopy images of KG1a cells perfused into the microfluidic chamber at the shear stress of 0.25 dyne cm⁻².
• Movie S2 (.mp4 format). Time-lapse transmitted light microscopy images of KG1a cells perfused into the microfluidic chamber at the shear stress of 0.5 dyne cm⁻².
• Movie S3 (.mp4 format). Time-lapse transmitted light microscopy images of KG1a cells perfused into the microfluidic chamber at the shear stress of 1.0 dyne cm⁻².
• Movie S4 (.mp4 format). Time-lapse transmitted light microscopy images of KG1a cells perfused into the microfluidic chamber at the shear stress of 2.0 dyne cm⁻².
• Movie S5 (.mp4 format). Time-lapse transmitted light microscopy images of KG1a cells perfused into the microfluidic chamber at the shear stress of 4.0 dyne cm⁻².
• Movie S6 (.mp4 format). Time-lapse transmitted light microscopy images of KG1a cells perfused into the microfluidic chamber in the presence of EDTA (10 mM) at the shear stress of 1.0 dyne cm⁻².
• Movie S7 (.mp4 format). Time-lapse transmitted light microscopy images of KG1a cells perfused into the microfluidic chamber at the shear stress of 1.0 dyne cm⁻².
• Movie S8 (.mp4 format). Time-lapse transmitted light microscopy images of MβCD-treated KG1a cells perfused into the microfluidic chamber at the shear stress of 1.0 dyne cm⁻².