Fig. 5: Large glycoproteins avoid positive membrane curvatures in vitro.

A Schematic illustration of supported lipid bilayers (SLB)-based arrays employed to study the in vitro curvature preference of a mucin-like glycoprotein, Podocalyxin (Podxl). B Cartoons elucidating the opposite membrane curvature for the distribution of mucin glycocalyx proteins on lipid bilayers vs. in the cell-based experiments. C Heatmaps show the spatial distributions of Podxl, deglycosylated Podxl, and lipid bilayers around the nanoXs (with 30% DGS-Ni-NTA). D, E Quantification of end-to-side (reflecting the preference for negative curvature) and inner-to-side ratios (reflecting the preference for positive curvature) of (D) Podxl and (E) deglycosylated Podxl on gradient nanoXs arrays of three selected inner angles. Podxl preferentially accumulates at negatively-curved membranes at the ends of nanoX arms. The lipid bilayers were doped with 30% DGS-Ni-NTA (see Supplementary Table 5A, B for the detailed statistics). F Heatmaps show the spatial distributions of Podxl, deglycosylated Podxl, and lipid bilayers around the nanoXs (with 10% DGS-Ni-NTA). G, H Quantification of end-to-side and inner-to-side ratios of (G) Podxl and (H) deglycosylated Podxl on the lipid bilayers doped with 10% DGS-Ni-NTA (see Supplementary Table 5C, D for the detailed statistics). All ratios are normalized against the rhodamine-lipid signals. Welch’s t-tests (unpaired, two-tailed, not assuming equal variance) are applied for all statistical analyses in this figure. Error bars represent SEM.