Figure 4.

Nef does not affect the cholesterol content of HIV-1 particles. (A) Quantitative lipid analysis was performed as described in Methods. Data are displayed as molar ratio of cholesterol to PC. Error bars represent standard deviation of the mean. (B) Selected high-field region of the¹H spectra of 0.4 mM Nef-SF2 (Δ_1–43, C210A) in the absence (black) and presence of 0.4 mM water soluble cholesterol (red), in comparison with water soluble cholesterol dissolved in the same buffer as the protein (green).(C) ¹H,¹⁵N-TROSY-HSQC spectra of 0.4 mM Nef-SF2 (Δ_1–43, C210A) highlighting an overlay of the C-terminal residues including the putative cholesterol binding site in the absence (black) and presence of 0.4 mM water soluble cholesterol (red) or 0.4 mM methyl-β-cyclodextrine (green).