Extended Data Fig. 10: Variable anti-PIP2 and anti-PIP3 antibody signal in synthetic condensates formed by LAF-1 RGG domains.

a, Metaplot of PIP2 and PIP3 signal proximal to condensates imaged in Fig. 6e. The location of condensates was identified in a semi-automated manner. Anti-PIP2 (above) or anti-PIP3 (below) immunofluorescence was then measured in the area surrounding each condensate’s center. The median GFP-LAF1 (red, left) and anti-PIP2 or anti-PIP3 (green, right) signal intensity across all examined synthetic condensates (from n = 3 biological replicates) is displayed with the distance from the center of each condensate indicated on the x-axis and y-axis. Median PIP2 and PIP3 immunofluorescence is higher in synthetic condensates formed by LAF-1 RGG domains than in adjacent regions. The number of condensates examined is indicated in each row. b, PIP2 and PIP3 are not enriched in synthetic condensates within select cells. A synthetic, condensate-forming protein containing GFP and two copies of the C. elegans LAF-1 protein’s RGG domain was over-expressed from a plasmid. Immunofluorescence was performed as described in Fig. 6a, using antibodies against either PIP2 (top) or PIP3 (bottom). Approximately two-third of the cells containing LAF-1 condensates had condensates that did not co-localize with either phosphoinositide and these images are representative of those cells. A region containing condensates is highlighted by a white square and expanded in the adjacent image (right). Scale bar, 5 μm (n = 3 biological replicates).