Figure 11. Both the OD and NBD of NPM1 are required for co-localization of rRNA and rpL5 within liquid-like droplets in vitro.
Confocal microscopic images 15 min after the addition of the specified NPM1 construct (30 μM) to preformed rpL5/rRNA puncta formed from 50 μg/mL rRNA and 200 μM rpL5 (a) and from 50 μg/mL rRNA and 50 μM rpL5 (b). Scale bar = 10 μm. Quantification of the growth of droplets formed through disassembly of rpL5/rRNA puncta upon addition of individual NPM1 constructs. (c) NPM1 constructs were titrated into puncta formed between 200 μM rpL5 and 50 μg/mL rRNA, where rpL5 is above the threshold for independent phase separation with NPM1. The star at 40 μM N294 indicates that the droplets experienced wetting on the slide surface and expanded above the maximum threshold of 100 μm2 set in the analysis; (d) NPM1 constructs titrated into puncta formed between 50 μM rpL5 and 50 μg/mL rRNA, where rpL5 is below the threshold for independent phase separation with NPM1.