Data analysis, functional annotation, and validation of potential nuclear speckle proteins. (A) Scatter plot of the nuclear speckle versus centromere ratio (SPK-CEN ratio) of proteins. (B) Bar chart showing the GO annotated biological processes of the top 250 proteins as sorted by the SPK-CEN-Ratio. (C) Pie chart showing the GO annotated biological processes of the top 100 proteins as sorted by the SPK-CEN ratio. (D) Immunofluorescence of U2OS cells costained with anti-MFAP1 or anti-ZNF207 (green) and SC35 (red) after PFA fixation. (E) Anti-SC35 immunostaining (red) of U2OS cells expressing GFP-ZNF207 (green) and fixed with PFA shows loss of GFP-ZNF207 fluorescence after fixation. Scale bars: 10 µm.