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. 2012 Mar 23;7(3):e34056. doi: 10.1371/journal.pone.0034056

Figure 5. Subcellular localization of BiFC signals.

Figure 5

αIIb-VCβ3 CHO cells were transduced with kindlin-2 (or control) shRNA lentiviruses also encoding DsRed, and VN-talin was induced by doxycycline. (A) Cells were incubated on fibrinogen-coated plates (100 µg/ml coating concentration) for 45 min, fixed, stained with antibody D57 for αIIbβ3, and examined by deconvolution microscopy (BiFC: Green, αIIbβ3: blue, Transduced: red). The arrows point to transduced cells, and the arrowhead to a non-transduced cell. (B) Spreading of transduced cells was examined and data were expressed as mean cell surface areas measured in total pixels as described in Experimental Procedures. Asterisk denotes statistically significant difference against respective control cells, P<0.01 (C) BiFC and αIIbβ3 fluorescence co-localization in transduced cells was evaluated by deconvolution microscopy as described in Experimental Procedures. Data represent 30–60 cells analyzed for each treatment. (D) Western blots were performed to monitor expression of talin and kindlin-2 in cell lysates. The cell lysates were from both uninfected and virus transduced cells whereas only virus transduced cells were analyzed in (A), (B) and (C).