Figure 4: Live cell imaging using tandem FP11 tag.
From: Versatile protein tagging in cells with split fluorescent protein
(a) Tracking of IFT particles in mouse IMCD3 cells expressing either IFT20::GFP or IFT20::GFP11x7+GFP1-10. Kymographs show both retrograde and anterograde transport (see Supplementary Fig. 3 and Supplementary Movie 1). (b) Comparison of IFT20 movement speed for IFT20::GFP and IFT20::GFP11x7. The number of particles that were analysed for each case is indicated in the figure. Error bars are standard deviations. (c) Comparison of the photobleaching rate in imaging β-tubulin labelled with GFP, GFP11 and GFP11x7, showing snapshots of S2 cells expressing the three different constructs at the beginning and the end of a 400 s movie. GFP11x7 samples were imaged with one-seventh the excitation laser power. Scale bars indicate 5 μm. (d) Fluorescence photobleaching time traces. Five cells were averaged in each condition. The error bars are standard deviations.
