Figure 2. Interaction of lifeAct with F‐actin and mutational analysis.

• A
  Surface representation of F‐actin bound to lifeAct as indicated.
• B
  Cartoon representation of lifeAct (red) in expanded view with n and n + 2^nd actin monomers (surface representation in magenta and blue), key interacting residues are highlighted.
• C
  Confocal images of U2OS cells transiently expressing lifeAct‐GFP wild‐type and mutants of lifeAct residues interacting with F‐actin, cells were additionally stained with SiR‐actin to confirm the actin filaments. The line scan as indicated with yellow line on the cells shows the extent of lifeAct (green) and SiR‐actin (magenta) co‐staining of actin structures. Scale bar = 5 μm.
• D
  Binding affinities calculated from titration data of co‐sedimentation assays of lifeAct‐14 (1.2 μM) and lifeAct‐17 (2.2 μM) as indicated. Data points for each concentration were averaged from three independent experiments; error bars represent SD between independent experiments.

Source data are available online for this figure.