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Figure 9


Fig. 9. Quantitative measurements of the size of the GFP-Bax complex and determination of the number of molecules within a single Bax complex at the MOM. (A) Image records of GFP-Bax small complexes in an apoptotic HeLa cell (left panel) and fluorescent beads (right panel) obtained under identical imaging conditions. Scale bar: 2 µm. (B) A sample image analysis of a 0.2 µm diameter bead using the MetaMorph software. (C) A calibration curve for converting the diameter from pixel numbers to µm based on the measurements of beads. (D) A sample image analysis of a GFP-Bax small complex. (E) Results of measurements on the size of GFP-Bax small complexes (n=316, from six cells). (F) Image record of purified GFP droplets obtained under identical imaging conditions as in A. Scale bar: 2 µm. (G) A calibration curve for converting the integrated fluorescent intensity to number of GFP molecules based on the measurements of GFP droplets. (H) Results of measurements on the number of GFP-Bax molecules within a Bax small aggregate (n=150, from six cells).