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Figure S4.
Further analysis of CMB proximity interactome, related toFig. 4. (A) Results of direct TurboID performed on CMB S2 cells (detergent-insoluble cytoskeletal fraction). LC-MS/MS analysis reveals centrosomal proteins (magenta), RNA-binding proteins (light green), and proteins involved in translation (dark green), chaperone-mediated protein folding (light blue), ubiquitination (blue), and proteolysis (dark blue). Volcano plots of −log10 P values against log2 fold change (sample/control). Significantly enriched proteins (log2 enrichment >1, P <0.05) are indicated in dark gray, with proteins of the above functional categories highlighted in color. See also Table S2 C. (B) Venn diagram revealing a significant overlap between CMB proximity interactome obtained from detergent-soluble (cytoplasmic) and detergent-insoluble (cytoskeletal) fractions of S2 cell extracts. See also Table S2 E. (C) Comparison of CMB S2 cell and testis TurboID interactomes with previous published datasets for centriolar satellites: the BioID of 22 satellite proteins mapped by Gheiratmand et al. (2019) and the PCM1-GFP pulldown performed by Quarantotti et al. (2019). Comparison of those proteins conserved between humans and flies. Numbers in parentheses are total number in each dataset. See also Table S2 E. (D) Comparison of BioID of 22 satellite proteins mapped by Gheiratmand et al. (2019) and PCM1-GFP pulldown performed by Quarantotti et al. (2019) with cytosolic RNA interactome defined by Youn et al. (2018). See also Table S2 F. (E) Single-molecule fluorescence hybridization (smFISH) combined with immunofluorescence microscopy shows Sas-4 mRNA colocalizing with nascent SAS-4 protein in the cytoplasm of S2 cells. (F and G) smFISH combined with immunofluorescence microscopy in S2 cells. Immunofluorescence micrographs (F) and corresponding quantitation (G). Sas-4 mRNA localizes in the vicinity of CMB foci. To exclude random colocalization/proximity, distribution was compared with randomized controls. N = 109 cells. Scale bars, 5 µm (E), 10 µm (F), 1 µm (E and F, insets).

Further analysis of CMB proximity interactome, related to Fig. 4 . (A) Results of direct TurboID performed on CMB S2 cells (detergent-insoluble cytoskeletal fraction). LC-MS/MS analysis reveals centrosomal proteins (magenta), RNA-binding proteins (light green), and proteins involved in translation (dark green), chaperone-mediated protein folding (light blue), ubiquitination (blue), and proteolysis (dark blue). Volcano plots of −log10 P values against log2 fold change (sample/control). Significantly enriched proteins (log2 enrichment >1, P <0.05) are indicated in dark gray, with proteins of the above functional categories highlighted in color. See also Table S2 C. (B) Venn diagram revealing a significant overlap between CMB proximity interactome obtained from detergent-soluble (cytoplasmic) and detergent-insoluble (cytoskeletal) fractions of S2 cell extracts. See also Table S2 E. (C) Comparison of CMB S2 cell and testis TurboID interactomes with previous published datasets for centriolar satellites: the BioID of 22 satellite proteins mapped by Gheiratmand et al. (2019) and the PCM1-GFP pulldown performed by Quarantotti et al. (2019). Comparison of those proteins conserved between humans and flies. Numbers in parentheses are total number in each dataset. See also Table S2 E. (D) Comparison of BioID of 22 satellite proteins mapped by Gheiratmand et al. (2019) and PCM1-GFP pulldown performed by Quarantotti et al. (2019) with cytosolic RNA interactome defined by Youn et al. (2018). See also Table S2 F. (E) Single-molecule fluorescence hybridization (smFISH) combined with immunofluorescence microscopy shows Sas-4 mRNA colocalizing with nascent SAS-4 protein in the cytoplasm of S2 cells. (F and G) smFISH combined with immunofluorescence microscopy in S2 cells. Immunofluorescence micrographs (F) and corresponding quantitation (G). Sas-4 mRNA localizes in the vicinity of CMB foci. To exclude random colocalization/proximity, distribution was compared with randomized controls. N = 109 cells. Scale bars, 5 µm (E), 10 µm (F), 1 µm (E and F, insets).

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