Supplementary Figure 4: Digitonin disrupts TCR nanocluster. | Nature Immunology

Supplementary Figure 4: Digitonin disrupts TCR nanocluster.

From: Inhibition of T cell receptor signaling by cholesterol sulfate, a naturally occurring derivative of membrane cholesterol

Supplementary Figure 4

(a) The purified TCRs were reconstituted in liposomes with indicated lipid compositions. The proteoliposomes were lysed with 1% digitonin and the lysates were subjected to Blue-Native PAGE and immune-blotted with anti-CD3ζ antibodies. The ferritin markers, f1 and f2 corresponding to 440 kDa and 880 kDa, are shown. Band intensities of nanoclustered and monomeric TCR were quantified with the Li-Cor Odyssey infrared imager and shown as a ratio of the two forms. Quantification data were collected from three independent experiments shown on the right.

(b) After DMSO or CS-treatment, the M.mζ-SBP cells were lysed with 1% digitonin. Purified protein complexes were subjected to Blue-Native PAGE and anti-CD3ζ immuno-blotting. Band intensities of nanoclustered and monomeric TCR were quantified with the Li-Cor Odyssey infrared imager and shown as a ratio of the two forms. Quantification data were collected from three independent experiments shown on the right.

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