Journal: Developmental cell

Article Title: PUF partner interactions at a conserved interface shape the RNA binding landscape and cell fate in Caenorhabditis elegans

doi: 10.1016/j.devcel.2024.01.005

Figure Lengend Snippet: (A) Tethering assay. FBF-2 is tethered by λN22 binding to boxB hairpins in the reporter RNA. Modified from Aoki et al. 201891. (B) Above, location of gonadal arm (black box) within animal. Below, location of distal region (dotted box) within gonadal arm. Asterisk marks distal end. Germline stem cells (GSCs) reside at the most distal end of germline and their daughters begin differentiation as they move proximally. (C) Diagram of distal gonad, showing distance from the distal end in microns below and extents of abundant FBF-2 (black line) and lower FBF-2 (dotted line) above. (D-E) Representative z-projections of extruded germlines. 20μm scale bar in (D) applies to all images. Dotted line marks gonad boundary; asterisk marks distal end. Top, GFP reporter expression (green); bottom, FBF-2FLAG staining (magenta). (D) Untethered wild-type FBF-2. (E) Tethered wild-type FBF-2. (F) ImageJ quantitation of reporter signal from tethered vs untethered FBF-2. GFP abundance plotted against distance from distal end. Solid line shows mean abundance and shading shows 95% confidence interval. Each plot represents three biological replicates with at least 10 gonad arms per replicate. P-values are given for pooled data in 0-35, 35-70 and 70-100 μm regions (black bars). P-values: *** p < 0.001, ** p < 0.01, * p < 0.05, ns (not significant) p > 0.05. Exact p-values in Table S4. (G-H) Representative z-projections of extruded germlines, as in (3D-E). (G) Untethered Y479A. (H) Tethered Y479A. (I) Quantitation of tethered vs untethered Y479A. P-values as in (F). (J) Western blots after FBF-2 and NTL-1 co-immunoprecipitation. Left, input lysates (1%); right, FLAG IP (10%). NTL-1:V5 co-immunoprecipitates with both wild-type and Y479A.

Article Snippet: mouse monoclonal αV5 , Bio-Rad , Cat# MCA1360, RRID:AB_322378.

Techniques: Binding Assay, Modification, Expressing, Staining, Quantitation Assay, Western Blot, Immunoprecipitation

Journal: Developmental cell

Article Title: PUF partner interactions at a conserved interface shape the RNA binding landscape and cell fate in Caenorhabditis elegans

doi: 10.1016/j.devcel.2024.01.005

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: mouse monoclonal αV5 , Bio-Rad , Cat# MCA1360, RRID:AB_322378.

Techniques: Virus, Recombinant, Protease Inhibitor, Software, Sequencing