Fig. 7: The RRM1 and RRM3 of Rim4 cooperate in recruiting mRNAs and Pab1 during SG growth.
From: Rim4-seeded stress granules connect temperature sensing to meiotic regulation
A–C FM of cells expressing Rim4 variants and mScarlet-Pab1. A Top: schematic depicts the mutation sites (red X) in GFP-Rim4(RRM3m). Bottom: Representative FM images. Yellow boxes and dashed lines are as described in Fig. 1F. B Average foci size analysis. N (foci) = 50. Red lines: median ± 95% CI. Unpaired two-sided Mann–Whitney test. p-values: a: 1.49e-8 (***); b: 7.59e-7 (***); c: 0.0021 (**); d: 0.0075 (**). C Percentage of Pab1/Rim4 (beige) and Rim4/Pab1 (brown) co-localization per cell. N.A.: not applicable (no mScarlet-Pab1 foci). Dots: cells analyzed. Red lines: mean ± SE. N (cells) = 50. Unpaired two-sided Mann–Whitney test. p-values: a: 1.98e-29 (***), b: 2.51e-10 (***); c: 0.3736 (ns); d: 0.9719 (ns); e: 0.0856 (ns); f: 1.98e-29 (***). D–G FISH visualizing Rim4 variants and mRNAs. D Top: schematic of the GFP-Rim4(NES-RRM1,3m). NESHst2 (Blue box): nuclear export signal derived from Hst2. Bottom: Representative FISH. Yellow boxes and dashed lines are as described in Fig. 1F. E Percentage of Rim4 foci co-localization with mRNAs, F foci sizes, and G Rim4 foci co-localization with mRNAs categorized by foci size, as described in Fig. 2D. E, F and small boxes in (G), N (foci) = 224, 102, 108; G red bars (S): foci size ≤ median, blue bars (L): foci size > median, N (foci) = 112, 112, 51, 51, 54, 54. (E) and bar graph of (G) data were shown as mean ± SE. (F), data were shown as median ± 95% CI (red lines). Small boxes in (G) were derived from (F), dashed lines: median. Data were analyzed by (E, F) unpaired two-sided Mann–Whitney test or (G) chi-square test. Two-sided p-values: E a: 1.70e-14 (***); b: 4.91e-6 (***); c: 0.0092 (**). F a: 9.34e-18 (***); b: 2.90e-10 (***); c: 0.0322 (*). G a: 0.0286 (*); b: 0.4898 (ns); c: 0.2453 (ns). H Model for the role of mRNAs in SG biogenesis. Scale bars are 5 µm.
