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. Author manuscript; available in PMC: 2016 Nov 23.
Published in final edited form as: Nat Immunol. 2016 May 23;17(7):844–850. doi: 10.1038/ni.3462

Figure 3. Cholesterol sulfate associates with TCR and disrupts cholesterol-TCR interaction.

Figure 3

(a) Mass spectrometry quantification of TCR- and TfR- associated CS with, CS (+supp), and without, CS (-supp), pre-incubation of the cells with CS. Data are presented as the ratio of CS amounts. (b) Immunoblot analysis of TCR in cholesterol beads treated with vehicle control, digitonin, CS or cholesterol. Band intensities of CD3ε were quantified with the Li-Cor Odyssey infrared imager and shown as arbitrary units (AU) in the right panel. (c) Autoradiogram and immunoblot analysis of TCRβ-cholesterol in the absence or presence of CS, then left untreated or subjected to PNGase F treatment as indicated. Proteins were separated by SDS-PAGE. The autoradiogram and the anti-CD3ε immunoblot are shown. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001; NS, not significant, unpaired t-test mean and s.e.m. (a,b). Data are from one experiment representative of three independent experiments with similar results (c) or three independent experiments with biological duplicates in each (a,b; n = 3).