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. 1999 May 11;96(10):5394–5399. doi: 10.1073/pnas.96.10.5394

Figure 4.

Figure 4

Flow cytometric analysis, dose-response, and competition curves of CHO DUKX-B11 cells labeled with fMTX. (A) Induced formation of FKBP–rapamycin–FRB complex was monitored by fluorescence flow cytometry. The gray histogram corresponds to cells expressing FRB–F[1,2] and FKBP–F[3] that had been treated overnight with 20 nM rapamycin. The white histogram corresponds to untreated cells. (B) The dose-response curve for rapamycin was based on flow cytometric analysis of CHO DUKX-B11 cells expressing the same fusions. Mean fluorescence intensities were determined for three independent samples at each rapamycin concentration (between 0.1 nM and 300 nM). (C) Competition curve with the inhibitor FK506, an analog of rapamycin. Mean fluorescence intensities were determined for three independent samples at each inhibitor concentration (between 0 μM and 6 μM, corresponding to a ratio of rapamycin:FK506 of 1:0 to 1:300; closed circles). The concentration of rapamycin was kept constant at 20 nM. As a control, mean fluorescence intensities also were determined for each concentration of FK506 in the absence of rapamycin (open triangles).