Figure 1

Exposure to 7-KC inhibits FRET between CD3 ε and Thy-1 molecules expressed on plasma membrane of CD4⁺T cells. FRET between Thy-1 and CD3ε was established using donor and acceptor pair of anti-CD3ε PE (FL2) and anti-Thy1 AlexaFluor 647 (FL3) in lymph node cells respectively. (A) To assess the FRET positive events the quadrant gates were set up in the absence of either the acceptor (control 1) or donor (control 2). Upper right quadrant shows FRET positive events when cells were stained with both the donor (anti-CD3ε) and acceptor (anti-Thy-1) fluorophore-conjugated antibodies with or without exposure to three different concentrations of 7-KC. (B) Representative images of control FRET experiment using anti-CD71 PE (FL2) as FRET donor and Anti-Thy1 AlexaFluor 647 (FL3) as acceptor. (C) Graphic representation of the number of events observed in A & B. A representative experiment of three independent replicates is shown.