Analysis of interdependence and colocalization of mRuby-PKR and PBs. (A) Representative immunofluorescence images showing two populations of mRuby-PKR clusters based on their diameter and association with Edc3. The image crop shows a close-up of these two populations. Scale bar: 10 µm. The right panel shows the quantification of the data (n = 30 cells). (B) Representative micrographs of fixed cells coexpressing mRuby-PKR and GFP-Dcp1a. The image crop shows two mRuby-PKR cluster populations, those that associate with GFP-Dcp1a and those that do not. Scale bar: 10 µm. (C) Representative immunofluorescence images showing pharmacological (CHX) or genetic (4E-T RNAi) knockdown of PBs assessed by Edc3 staining. Scale bar: 10 µm. (D) Quantification of the data in C. (N = 3 experiments, n > 500; ****, P < 0.0001, unpaired Student’s t test, nonparametric). (E) Representative micrographs showing that mRuby-PKR clusters recruit 4E-T after pharmacologic depletion (CHX) of PBs. Scale bar: 10 µm. (F) Western blot showing the extent of knockdown of 4E-T KD by RNAi. Right: Quantification of the data (mean and SEM, N = 3 experiments; ****, P < 0.0001, unpaired Student’s t test, nonparametric). Source data are available for this figure: SourceData FS2.