Cholesterol sequestration prevents the induction of uropod structures in T lymphoblasts.

Control (A,B) or T lymphoblasts stimulated with 100 ng/mL SDF-α (C,D) or 1 μg/mL anti–ICAM-3 HP2/19 mAb (E-G) were treated (B,D,F,G) or not (A,C,E) with 8 mM CD in RPMI medium. After CD treatment, part of the cell culture stimulated with anti–ICAM-3 HP2/19 mAb was washed and incubated in RPMI medium supplemented with 20% FCS in the absence of CD (G). Bar is 20 μm. Panel H shows the cholesterol levels, as detected by staining with filipin, in control (−) or in T lymphoblasts treated with CD. Bright field images are depicted to show the cell field. The presence of uropod in approximately 500 cells was analyzed in 3 independent replicates. The results obtained in untreated (gray bars) and CD-treated (black bars) cells are represented as percentage of uropod-bearing cells present in the culture (I). The mean ± SD is shown.