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Fig. 4. Cellular cholesterol levels affect chemotactic cell migration. (A) CD24+/+ 18H18+ cells were loaded with cholesterol by incubation with 30 µg/ml soluble cholesterol and tested for chemotactic cell migration in response to SDF-1. CD24–/– cells N232.18 and 18H18– were treated with fluvastatin for 72 hours and then tested for chemotactic cell migration in response to SDF-1. Data are presented as the percentage migration (± s.e.m.) compared with untreated control cells. (B) SDF-1-triggered ERK-phosphorylation in CD24–/– (N232.18 and 18H18–) and CD24+/+ (18H18+) cells. Cells were exposed for the indicated length of time (in minutes) to 100 ng/ml SDF-1 then lysed. An equal volume of lysate was mixed with sample buffer and analysed by SDS-PAGE and western blotting using a phospho-ERK-specific antibody and ECL detection. For a loading control, blots were reprobed with an ERK-1-specific antibody. (C) CD24–/– (18H18–) cells were treated with fluvastatin for 72 hours and then ERK phosphorylation in response to SDF-1 was tested. Note that ERK phosphorylation in 18H18– cells was abolished after fluvastatin treatment.





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