Fig 4. Cigarette smoke increases caveolin mediated endocytosis.

Primary human SAEC were exposed to either air or cigarette smoke for 60 minutes at the air-liquid interface. (A) Following exposure cells were rested overnight and treated with AF594-labeled CtxB (10μg/mL) for 30 min and fixed for confocal microscopy. or BODIPY-LacCer (1μM) as described in the materials and methods. (B) Percentage of cells taking up AF594-CtxB was quantified as described in the Methods, using 3 high power fields per culture and three replicate cultures of one SAEC cell strain. (C) SAEC exposed to air or smoke were rested for 5 hr then treated with AF488-CtxB (10μg/ml) as described in materials and methods and were analyzed by flow cytometry. Each pair of points represents an SAEC strain from a different donor. ***p<0.001 by two way ANOVA with Sidak correction. (D) Photographs of air and smoke-exposed cells treated with BODIPY-LacCer.