GM1 levels and Thy-1 GPI-anchored protein expression are higher in lipid raft domains from CD8+ than CD4+ T cells. (A) Splenic T cells from young mice were treated with Triton X-100 and lysates were submitted to ultracentrifugation on a sucrose gradient for lipid rafts isolation. Equal volumes of fractions were analyzed by dot blot analysis using CTB subunit conjugated to HRP. Raft and non-raft fractions are indicated. (B) Raft or non-raft fractions of CD4 and CD8 cell lines were pooled together and immunoprecipitated with mAb specific for Thy1 and subsequently detected using anti-thy1 antibody. (C) The relative density of thy1 bands seen in (B) for CD4+ and CD8+ cells were calculated using the Image Quant software (Amersham Biosciences) and shown in bar diagram. Black bars represent raft-pooled fractions and white bars, non-raft pooled fractions.