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Fig. 8 Spplement Cholesterol mediates DIM-488 spread on A549 epithelial cells. (A) Structure of alkyne-cholesterol. (B) Structure of 16:0 azidocaproyl phosphoethanolamine (Azido-PE). (C) Images of wildtype (WT) and ∆mmpL7 M. marinum expressing a green-fluorescent protein following 48 h incubation with azido-PE reacted with DIBO-647, scale bar = 3 μm. (D) Mean (± SEM) fluorescent intensity of line profiles drawn perpendicular to poles of WT and ∆mmpL7 M. marinum labeled as in C. Representative of two separate experiments. (E) Mean cholesterol content of A549 cells treated with methyl-ß cyclodextrin (MßCD), water-soluble cholesterol, or both. (F) Mean percent DIM-488 spread at 24 hpi of A549 epithelial cells treated with MßCD, water-soluble cholesterol, or both, MOI = 5. (E) and (F) representative of three separate experiments.