Journal: Nature

Article Title: Probing condensate microenvironments with a micropeptide killswitch

doi: 10.1038/s41586-025-09141-5

Figure Lengend Snippet: a , Schematic of the NuFANCI procedure. b , Protein expression profiles of NuFANCI-isolated nucleoli and their respective input samples. In the left four columns, nucleolar proteins are annotated based on previous proteomics data. N, two clusters of nucleolar proteins. LFQ, label-free quantification. c , The proteomes of GFP-nb–KS-targeted nucleoli and GFP-nb–KS F-to-G -targeted nucleoli compared with the anti-GFP-nanobody control. P values were calculated using one-sided Student’s t -tests. FC, fold change. d , Fixed-cell immunofluorescence images of GFP–NPM1-expressing cells that were untransfected or transfected with GFP-nb, GFP-nb–KS, GFP-nb–2×KS or GFP-nb–KS F-to-G constructs. Scale bars, 5 µm. e , NEPRO fluorescence intensity in nucleoli. Data are mean ± s.d. P values were calculated using one-way ANOVA followed by Dunnett’s T3 multiple-comparison test versus the GFP-nb condition. n = 32 (untransfected), 33 (GFP-nb), 36 (GFP-nb–KS), 29 (GFP-nb–2×KS) and 31 (GFP-nb–KS F-to-G ) cells from two independent experiments. f , NEPRO fluorescence intensity in nuclei. Data are mean ± s.d. P values were calculated using one-way ANOVA followed by Dunnett’s T3 multiple-comparison test versus the GFP-nb condition. The numbers of cells are the same as described in e and are from two independent experiments. g , FRAP analysis of mCherry-signal in HCT-116 cells expressing GFP–NPM1 from the endogenous locus after transfection with the indicated nb constructs and co-transfection with mCherry–SURF6 or mCherry–RPL18. Data are mean ± s.d. n = 10 for all, except for the SURF6 experiment for the TagBFP-only sample ( n = 8; asterisk). h , Fixed-cell immunofluorescence images of 5.8S rRNA in GFP–NPM1-expressing cells that were transfected with either the GFP-nb or GFP-nb–2×KS construct. Scale bars, 5 µm. i , Quantification of 5.8S rRNA mean fluorescence intensities in the demixed and remaining portions of nucleoli in GFP-nb–2×KS-expressing cells. Data are mean ± s.d. n represents the number of cells from one experiment. The experiment was repeated twice with similar results. P values were calculated using one-way ANOVA followed by Tukey’s post hoc test. Some elements in the scheamtic in a were created in BioRender. Hnisz, D. (2025) https://BioRender.com/fa5zmne .

Article Snippet: The following primary antibodies were used: 5.8S rRNA (Novus, NB100-662SS, 1:500), HA-tag (Cell Signaling, C29F4, 1:1,000), NEPRO (Santa Cruz, sc-376579, 1:100), RNAPII (Abcam, ab26721, 1:500) and H3K27Ac (Abcam, ab4729, 1:1,000).

Techniques: Expressing, Isolation, Quantitative Proteomics, Control, Immunofluorescence, Transfection, Construct, Fluorescence, Comparison, Cotransfection

Journal: PLoS ONE

Article Title: Time-Dependent Decay of mRNA and Ribosomal RNA during Platelet Aging and Its Correlation with Translation Activity

doi: 10.1371/journal.pone.0148064

Figure Lengend Snippet: On day 8, washed PLTs from DT- or saline-treated mice were resuspended in Tyrode’s albumin/DMEM medium and incubated at 37°C for different periods of time. (A) PLTs were incubated for 0, 2, 4, 6 and 24 h. The percentage of TO bright PLTs, determined at 0, 6 and 24 h, was 82, 33 and 2%, respectively. At each time points, PLT samples were fixed and permeabilized, stained with the anti-rRNA mAb Y10b and A647-conjugated anti-mouse IgGs, counterstained with an A488-conjugated anti-GP1bβ mAb and analyzed by confocal microscopy. PLT outlines, determined by Photoshop analysis of anti-GP1bβ staining, are depicted. Representative micrographs of analyzed cells from DT-treated animals (+DT) at times 0, 4 and 24 h and from saline-treated animals (-DT) at time 0 are shown. (B) The mean fluorescence intensity per unit area in each PLT stained with Y10b (+) or an isotype-matched control antibody (C) (y-axis, log2 scale) was calculated as described in the methods; 500 to 550 PLTs from DT-treated mice (+) or saline-treated mice (-) were analyzed for each condition. A one way ANOVA analysis followed by a Bonferroni post-hoc test comparing each pair of conditions gave p values of <0.001 (continuous bars) or <0.01 (dashed bars). Other combinations of Y10b stained samples were not statistically different.

Article Snippet: The anti-rRNA mAb Y10b (5 μg/mL, mouse IgG3, Novus Biologicals) was used to label ribosomes and revealed with A647-conjugated goat anti-mouse IgG.

Techniques: Saline, Incubation, Staining, Confocal Microscopy, Fluorescence, Control