(A) Summary of physical properties and phase separation behavior of selected sterols. Topological Polar Surface Area (TPSA) computed surface sum over all polar atoms. Ordered lipid (Lo) domain (raft) stabilizing (+) or disrupting (−) sterols are indicated relative to cholesterol (++). Refer to Table S2 for additional details.
(B) Sterol profiles of AY9944-treated HEK293T cells after sterol loading (mean ± SD). n = 4 independent biological replicates from 2 MβCD-sterol preparations.
(C) Tfn uptake relative to LPDS controls (mean ± SD). One-way ANOVA (F(12, 51) = 11.23, p < 0.0001) and Dunnett’s test versus LPDS control (n = 5 biological replicates from 3 independent experiments, ~1,500 cells per replicate). (c), commercially available MβCD-Chol.
(D) Representative mid-plane confocal images after incubation with AF-555 conjugated Tfn. Scale bar, 20 μm.