Fig. 4 |. Edc3 couples activation of decapping to phase separation.

a, Schematic of in vitro decapping assay to determine contribution of decapping activity from Dcp1/Dcp2_ext in droplets (Dense Phase) and outside droplets (Dilute Phase). b, Dcp1/Dcp2_ext localized in droplets does not strongly contribute to overall activity (blue bars) but removal of Dcp1/Dcp2_ext/Edc3 droplets strongly diminishes activity (pink bars). Concentration of Dcp1/Dcp2_ext is 100 μM and 5 μM in absence pr presence of 80 μM Edc3, respectively. Relative activity refers to ratio of observed rate to dilute phase. c, Activation of decapping by Edc3 is concomitant with formation of microscopically visible droplets. Removal of droplets abrogates activation by Edc3. d, Edc3 activation coincides with depletion of Dcp1/Dcp2_ext (red squares) from solution. Curve showing mean Edc3 activation is reproduced from c for purposes of comparison. Relative activity refers to ratio of observed rate to Dcp1/Dcp2_ext in dilute phase without Edc3. Decapping data are from two independent experiments. Pelleting data are presented as mean ± s.e.m. for three independent experiments. Representative micrographs are from three independent experiments with similar results.