Figure 3. G3BP1 and IMP1 are enriched in... | Rockefeller University Press

Figure 3.

$Figure 3. G3BP1 and IMP1 are enriched in distributed nanocores within SGs. (A) Single-molecule imaging of SiR-labeled SNAP-G3BP1 and TMR-labeled HaloTag-IMP1. Snapshots before image acquisition confirmed the presence of SGs and showed that G3BP1 and IMP1 colocalized in the same granules (left). The outline of the cell and the nucleus are indicated. Localizations of single molecules in SGs of the indicated region (yellow box in the snapshot images) are shown (right). Bars: (left, middle) 10 µm; (right) 1 µm. (B) Spatial clustering of single-molecule localizations of an area within a single SG after combined expression of G3BP1 and IMP1. Raw localizations (left), the respective cluster patterns (middle), and cluster pattern overlap (right) are shown. (C) Quantitation of the fraction of the granular area, which is occupied by clusters, and weighted overlap for different combinations of expressed RBPs. The value for “n” represents the number of individual granules analyzed. Granules were obtained from 5–19 cells from three to eight independent experiments. For all experiments, stress had been induced by a 20-min treatment with 0.5 mM sodium arsenite before imaging.$

G3BP1 and IMP1 are enriched in distributed nanocores within SGs. (A) Single-molecule imaging of SiR-labeled SNAP-G3BP1 and TMR-labeled HaloTag-IMP1. Snapshots before image acquisition confirmed the presence of SGs and showed that G3BP1 and IMP1 colocalized in the same granules (left). The outline of the cell and the nucleus are indicated. Localizations of single molecules in SGs of the indicated region (yellow box in the snapshot images) are shown (right). Bars: (left, middle) 10 µm; (right) 1 µm. (B) Spatial clustering of single-molecule localizations of an area within a single SG after combined expression of G3BP1 and IMP1. Raw localizations (left), the respective cluster patterns (middle), and cluster pattern overlap (right) are shown. (C) Quantitation of the fraction of the granular area, which is occupied by clusters, and weighted overlap for different combinations of expressed RBPs. The value for “n” represents the number of individual granules analyzed. Granules were obtained from 5–19 cells from three to eight independent experiments. For all experiments, stress had been induced by a 20-min treatment with 0.5 mM sodium arsenite before imaging.