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. 2020 Apr 27;9:e56731. doi: 10.7554/eLife.56731

Figure 7. SIMR-1 localizes to foci adjacent to P granules and Mutator foci.

(A) Immunostaining of SIMR-1 (green) with MUT-16 (red, top left), PGL-1 (red, top right), ZNFX-1 (red, bottom left), and RSD-2 (red, bottom right) demonstrates that SIMR-1 localizes to foci near Mutator foci (MUT-16), P granules (PGL-1), and Z granules (ZNFX-1) but overlaps most substantially with RSD-2 foci. Arrow indicates an example of a single Z granule associated with two SIMR-1 foci. (B) Bar graph showing distance between the centers of fluorescence for indicated proteins to SIMR-1 (mean +/- SEM). See Materials and Methods for description of quantification methods. n.s. denotes not significant and indicates a p-value>0.05, * indicates a p-value≤0.05, ** indicates a p-value≤0.01, **** indicates a p-value≤0.0001. See Supplementary file 8 for more details regarding statistical analysis. (C) Immunostaining of SIMR-1 (green), ZNFX-1 (red), and PGL-1 (white) allows for visualization of the stacked SIMR/Z granule/P granule foci. All images are projections of 3D images following deconvolution. DAPI is blue in all panels and scale bars are 5 μm.

Figure 7—source data 1. Data used to generate Figure 7B.

Figure 7.

Figure 7—figure supplement 1. SIMR-1 and PRG-1 localize independently and to distinct granules.

Figure 7—figure supplement 1.

(A) Immunostaining of SIMR-1::GFP (green) and PRG-1::mKate (red) demonstrates that SIMR-1 and PRG-1 localize near one another but their localization is not coincident. (B) Live imaging of SIMR-1::GFP in a prg-1 mutant (top) and PRG-1::mKate in a simr-1 mutant (bottom) indicate that prg-1 is not required for SIMR-1 foci formation, nor is simr-1 required for PRG-1 localization to P granules. (C) Western blot of mKate2::3xMyc::PRG-1 in a wild-type or simr-1 mutant background demonstrate that the simr-1 mutation does not affect PRG-1 protein expression. Actin protein is shown as a loading control. Scale bars, 5 μm.