Client recruitment in vitro. (A and B) CGB-GFP (2.5 µM; green) was mixed with Cy3-tagged LyzC (1 µM; red) and was subsequently followed by addition of 20 mM calcium to induce liquid-like droplets or with 20 mM zinc to induce aggregates. While calcium-induced CGB-GFP droplets (top) recruit Cy3-LyzC, the zinc-induced aggregates (bottom) fail to recruit it. Bar graph is B quantifies the ratio of intensities in Cy3 v/s GFP channel, imaged at the same acquisition settings, to monitor recruitment of LyzC in relation to amount of CGB in the droplet (red) v/s aggregate (blue). Data is represented as mean ± SD from 28 droplets and 27 regions within aggregates. Statistical analysis was performed by unpaired two-tailed t test. ***P < 0.001. (C) CGB-GFP (2.5 µM; green) was mixed with Cy3-tagged CatD (1 µM; red) and droplet formation was induced using 3% PEG 8000 to monitor the recruitment of clients into the condensates. Note that Cy3-CatD shows strong recruitment to the droplets.