FIGURE 5

Secretome analysis of WT, COG4^p.G516R, and COG4-KO chondrosarcoma cells. (A) Coomassie blue staining of proteins in cell lysates and conditioned medium from COG4^p.G516R clones #1, #2, WT controls and COG4 KO cells. (B) Volcano plot of differentially secreted proteins between COG4^p.G516R and WT. (C) Volcano plot of differentially secreted proteins between COG4^p.G516R and COG4-KO cells. A few top candidates confirmed by pathway analysis were labeled respectively. (D) Dot plot showing enriched pathways from Gene Set Enrichment Analysis with the Normalized Enrichment Score (NES) shown on the X-axis. The size of the dots represents the number of genes in the significant Data Set list associated with the GO term and the color of the dots represent the adjusted p- values. (E) Table showing a list of top 10 candidates which were decreased in secretions of COG4^p.G516R with log 2 (Fold Change) and adjusted p-value shown for COG4^p.G516R vs. WT and COG4^p.G516R vs. COG4-KO. The number of peptides is equal to or more than two for all the candidates shown in table. (F) Western blotting of top candidates in medium decreased in both COG4^p.G516R and COG4-KO cells compared to WT. GAPDH was from cell lysates with comparable amount as loading control. (G) Western blotting of top candidates only changed in COG4^p.G516R compared to WT and COG4-KO cells. All experiments were performed in duplicate with similar results.