Fig 1: Study of the YFP-SPANX-A interactome A375 human melanoma cell line. (A) Gene Ontology based on the biological function and molecular processes of co-precipitated proteins together with SPANX-A in A375 melanoma cell line. The ten most enriched processes are shown. P < 0.05 (B) Representative scheme of principal putative interactors of SPANX-A based on their function. Information was obtained from Uniprot Database. Only the most representative functions are shown. Only the 19 most intense ribosomal proteins are shown (C) Validation of the interaction between YFP-SPANX-A and Lamin A/C by co-immunoprecipitation and western Blotting (N = 3). Crop from Supplementary Fig. S4. (D) Validation of the co-localisation between YFP-SPANX-A and Lamin A/C by immunofluorescence. YFP-SPANX-A is represented in green and Lamin A/C is shown in red. For the specificity of the secondary antiserum, the primary antibody was omitted. The nuclei were stained with Hoechst and are represented in blue. Scale bar 15 µm. (N = 3) Images were evaluated using ImageJ software (1.48v) (E) Co-localisation between YFP-SPANX-A and HSPA8, and YFP-SPANX-A and RPL38 by immunofluorescence. For the specificity of the secondary antiserum, the primary antibody was omitted. The nuclei were stained with Hoechst and are represented in blue. Scale bar 15 µm. (N = 3) Images were evaluated using ImageJ software (1.48v) (F) Expression of Lamin A/C and HSPA8 in A375SPANX-KD cells. A375SPANX-KD: Stable SPANX-A/D knockdown variant of the A375 human melanoma cell line; A375CTRL: control cell line. N = 3.
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