Figure S4. Characterization of the Inhibition of MEX-5 on mRNA-Dependent PGL-3 Drop Assembly, Related toFigure 4
(A) Effect of MBP on mRNA-dependent PGL-3 drop assembly. Plot of the fraction of total GFP fluorescence found in phase-separated drops of PGL-3-mEGFP
(0.6mM) on addition of 50 ng/mL mouse brain mRNA, in presence or absence of 150 nM MBP. The values were normalized to the case in absence of MBP. In each
case, drops inR 19 observation volumes (41mm 341 mm 3 10 mm) were scored. Error bars represent 1 SEM among the observation volumes scored.
(B) MEX-5 inhibits PGL-3 drop assembly dependent on mRNA purified fromC. elegans. Maximum intensity projections of series of confocal z- slices show drops
of PGL-3-mEGFP (0.6mM) that form on addition of 1 ng/mL or 2 nM totalC. elegansmRNA in presence or absence of 150 nM MBP-MEX-5 (236-350).
(C) Binding of PGL-3-mEGFP and MBP-MEX-5 (236-350) assayed in pull-down assay using Dynabeads Protein G beads coated with anti-PGL-3 antibody. SDS-
PAGE shows input, supernatant, and pellet fractions in Control: MBP-MEX-5 (236-350) alone (1mM), Experiment: MBP-MEX-5 (236-350) (1mM) and PGL-3-
mEGFP (1mM).
amelia
(Amelia)
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