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. 2014 Mar;34(5):862–876. doi: 10.1128/MCB.01227-13

FIG 3.

FIG 3

H-Ras expression inhibits K-Ras coclustering with PS on the PM. BHK cells expressing GFP–K-RasG12V alone (GFP-KG12V), GFP–K-RasG12V and RFP–K-RasG12V (GFP/RFP-KG12V), or GFP–K-RasG12V, RFP–K-RasG12V, and HA–H-RasG12V (GFP/RFP+HA-HG12V) were imaged in a FLIM. Representative images (a) and mean GFP lifetimes (calculated per cell) ± SEM (n ≥ 60 cells from 3 independent experiments) (b) are shown. BHK cells expressing GFP-LactC2 alone (GFP-LactC2), GFP-LactC2 and RFP–K-RasG12V (GFP-C2/RFP-Ras), or GFP-LactC2, RFP–K-RasG12V, and HA-tagged H-RasG12V (GFP/RFP+HA-HG12V) were imaged in a FLIM. Representative images (c) and mean GFP lifetimes (calculated per cell) ± SEM (n ≥ 60 cells from 3 independent experiments) (d) are shown. Control BHK cells (TF-PS) and BHK cells expressing RFP–K-RasG12V or coexpressing RFP–K-RasG12V and HA–H-RasG12V were flash-labeled with TF-PS and imaged in a FLIM. Representative images (e) and mean TF-PS lifetimes (calculated per cell) ± SEM (n ≥ 60 cells from 3 independent experiments) (f) are shown. The statistical significance of differences between mean lifetime values was evaluated using one-way ANOVA (*, P < 0.01).