Figure 2
From: Development of a quantitative method to measure EV uptake
DSP1 and DSP2 in vitro re-association assessment. (A) DSP1 and DSP2 reassociation kinetics in cell lysates. SUM159 stably expressing DSP1, DSP1-CD9 or DSP1-CD63 were lysed and each lysate was mixed with a cell lysate from DSP2-expressing cells. Renilla luciferase activity was measured with Stop&Glo Renilla reagent at the indicated times. n = 2. (B) Comparison of the signal obtained with Stop&Glo reagent at 24 h from combinations of cell lysates of cell expressing DSP1 constructs with cell lysates from cells expressing DSP2 (DSP1 + DSP2, DSP1-CD9 + DSP2 and DSP1-CD63 + DSP2) and with cell lysates from cells that co-expressed both constructs (DSP1/DSP2, DSP1-CD9/DSP2 and DSP1-CD63/DSP2). Negative controls of independent lysates from cells expressing either DSP1, DSP2, DSP1-CD9 or DSP1-CD63 are depicted. (C) Reassociation kinetics with EV lysates. EVs were isolated from conditioned media of SUM159 stably expressing DSP1, DSP1-CD9 or DSP1-CD63 by ultracentrifugation at 100000 g, lysed and each lysate was mixed with a cell lysate from DSP2-expressing cells. Renilla luciferase activity was measured with Stop&GloRenilla reagent at the indicated times.
