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. Author manuscript; available in PMC: 2013 Dec 21.
Published in final edited form as: ACS Chem Biol. 2012 Sep 25;7(12):1950–1955. doi: 10.1021/cb300280k

Figure 2.

Figure 2

Localization and translocation of PH(FAPP1)-mCherry-FKBP-Tiam1 at the Golgi (a) Schematic representation of the various FKBP-fused Tiam1 constructs (MFT and PMFT), control construct (MF), and plasma membrane tethered Lyn-GFP-FRB (LGR) construct. (b) Confocal fluorescent images showing the cellular localization of (i) MF, (ii) MFT, or (iii) PMFT with LGR at defined time points before (−) and after (+) rapamycin addition. Graph at the bottom of each column is a line scan of the respective cells prior to rapamycin addition (from respective red and green lines drawn on top panel cell images). Note that the LGR signal intensities are comparable in (i)– (iii). Scale bar beneath each graph represents 10 μm. (c) Average translocation kinetics of (i) MF, (ii) MFT or (iii) PMFT from the cytosol or Golgi (note y-axis), where light blue regions indicate presence of rapamycin. Signal intensities have been normalized against the maximum signal intensity values in each case. The average duration for MF, MFT and PMFT to reach half-maximal translocation were 11.9 s, 17.5 s and 27.9 s, respectively. S.E.M. values are indicated by error bars.

SC Phua et al.