Figure 5. G3BP1 and IMP1 exhibit a... | Rockefeller University Press

Figure 5.

$Figure 5. G3BP1 and IMP1 exhibit a biphasic partition in a bound and mobile fraction within SGs. (A) Example of a representative granule in a double-transfected cell. The granule was identified based on SiR staining (left) and localizations and trajectories based on TMR staining of HaloTag-G3BP1 (middle and right) are shown. The border of the SG is indicated by a dashed line. Bars, 1 µm. (B) Trajectory of a single G3BP1 molecule (as indicated by an arrowhead at time point 0) showing alternating phases of fast movement and immobile time periods. The trajectory is indicated in green. Bar, 250 nm. (C) High-mobility trajectories (mobile fraction, left) and low mobility trajectories (bound fraction, right) of HaloTag-G3BP1 within the granule shown in A. Bar, 1 µm. (D) Probability density of single-molecule diffusion coefficients from trajectories of a single granule expressing HaloTag-G3BP1 showing a biphasic partition in a bound (red) and mobile fraction (green). (E) Quantification of bound fractions of the respective Halo-tagged constructs expressed in different combinations. Calculations were performed on the total trajectories of 18–28 SGs per experimental condition as indicated by “n.” SGs were obtained from 12–15 cells from 3–10 independent experiments. For all experiments, stress had been induced by a 20-min treatment with 0.5 mM sodium arsenite before imaging.$

G3BP1 and IMP1 exhibit a biphasic partition in a bound and mobile fraction within SGs. (A) Example of a representative granule in a double-transfected cell. The granule was identified based on SiR staining (left) and localizations and trajectories based on TMR staining of HaloTag-G3BP1 (middle and right) are shown. The border of the SG is indicated by a dashed line. Bars, 1 µm. (B) Trajectory of a single G3BP1 molecule (as indicated by an arrowhead at time point 0) showing alternating phases of fast movement and immobile time periods. The trajectory is indicated in green. Bar, 250 nm. (C) High-mobility trajectories (mobile fraction, left) and low mobility trajectories (bound fraction, right) of HaloTag-G3BP1 within the granule shown in A. Bar, 1 µm. (D) Probability density of single-molecule diffusion coefficients from trajectories of a single granule expressing HaloTag-G3BP1 showing a biphasic partition in a bound (red) and mobile fraction (green). (E) Quantification of bound fractions of the respective Halo-tagged constructs expressed in different combinations. Calculations were performed on the total trajectories of 18–28 SGs per experimental condition as indicated by “n.” SGs were obtained from 12–15 cells from 3–10 independent experiments. For all experiments, stress had been induced by a 20-min treatment with 0.5 mM sodium arsenite before imaging.