Extended Data Fig. 5: NuFANCI proteome analysis of nucleoli targeted with killswitch.
From: Probing condensate microenvironments with a micropeptide killswitch
a. Principal Component (PC) analysis of NuFANCI-isolated nucleoli from untransfected cells, ActD-treated, GFP-nb, GFP-nb-KS, GFP-nb-2xKS, GFP-nb-KSF-to-G, and their respective input samples. The expression values were then standardized. b. Principal component analysis of NuFANCI-isolated nucleoli without input and ActD-treated samples. c. Correlation plot of NuFANCI samples using Pearson’s correlation coefficient. d. Euler diagrams showing the overlap of NuFANCI-detected proteins in nucleolus reference proteomes. e. Heatmap showing the protein expression profiles of NuFANCI-isolated nucleoli. Protein markers used in nucleolus isolation, mCherry and msfGFP-NPM1, some common nucleolar proteins, and protein marker (NEPRO) tested by IF are annotated on the right. Proteins have also been annotated based on the proteome of isolated nucleoli from previous proteomics data on the left. f. Barplots showing the percentage of proteins in the pooled input (top) or pooled nuFANCI samples (bottom) mapping to individual compartments annotated in the SubCellBarcode database. g. Quantification of NEPRO, SURF6, and RPL18 expression in NuFANCI samples shown in b. Data are mean ± s.d. n = 3 biologically independent experiments. h. Volcano plots of NuFANCI pairwise-comparisons. Significance values are from one-sided Student’s t-tests. Non-significant proteins are shaded in grey, whereas enriched and depleted are shaded in red and blue, respectively. Protein marker NEPRO was tested by IF.
