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. 2017 Jun 21;199(3):874–884. doi: 10.4049/jimmunol.1700431

FIGURE 1.

FIGURE 1.

Cav1 is localized within soluble and insoluble membrane fractions and regulates cholesterol and lipid composition. (A) Cav1 and Cav2 protein expression was compared between WT and Cav1-KO CD8 T cell lysates by Western blot with anti-Cav1 and Cav2 Ab. Cell lysates were immunoprecipitated with anti-Cav1 and Western blots were probed sequentially with anti-Cav1 and Cav2 Ab. (B) Cell lysates were separated into subcellular fractions (C, cytoplasmic; MS, membrane soluble; NS, nuclear soluble; CB, chromatin bound; LCC, lipid-cytoskeletal complexes) and analyzed by Western blot with anti-Cav1 Ab. (C) Total sphingomyelin content in WT versus Cav1-KO CD8 T cells. (D) Sphingomyelin species 14:0 and 16:0 are represented as a percentage of total sphingomyelin. (E) Free cholesterol from WT and Cav1-KO CD8 T cells is representative of one of three independent experiments from a total of 12 biological samples of each genotype. (F) Total ceramide and (G) ceramide monohexamide content. Lipidomics data are pooled from three biological repeats. Data are shown as mean + SD. *p ≤ 0.05, **p ≤ 0.01 (Student t test). ns, not significant.