Functionality of smORFs Venn diagram showing the number of smORF proteins identified by MS in different experiments. Functionality of smORFs Venn diagram.

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Functionality of smORFs Venn diagram showing the number of smORF proteins identified by MS in different experiments. Functionality of smORFs Venn diagram showing the number of smORF proteins identified by MS in different experiments. SDS gels: smORFs identified in SDS gels (n = 4); solution: smORFs identified from total protein extracts (n = 4); SEC_MS: smORFs identified in size‐exclusion chromatography (n = 2); DNAC_MS: smORFs identified in DNA–cellulose columns (n = 3). The graph represents the increase in the number of detected new smORFs (in orange) and known smORFs (in blue) by using different MS approaches. The names of MS approaches are as follows: Tris‐Gly (n = 2) and Bis‐Tis (n = 2) indicate smORFs identified by different SDS gels; solution: smORFs identified from total protein extracts (n = 4); SEC_MS1: smORFs identified in the first experiment of size‐exclusion chromatography (n = 1) and SEC_MS2: smORFs identified in the second experiment of size‐exclusion chromatography (n = 1); DNAC_RNA, DNAC_C and DNAC_NaCl: smORFs identified in DNA–cellulose columns, eluted by using RNA, chromatin or NaCl, respectively (n = 3); and sucrose indicates the number of smORFs identified by sucrose cushion. The graph represents the size‐exclusion chromatography elution profiles of different proteins: MPN516 (rpoB, subunit of the RNA polymerase complex), MPN606 (enolase, 50 kDa) and MPN476 (cytidylate kinase, 24 kDa). The main elution peak for the identified smORFs is indicated by dots. MPN155a and MPN060a were found after overexpression as a TAP fusion. The histogram represents the percentages of proteins in each category that have been identified to interact with DNA in affinity chromatography after receiving operating characteristic curve (ROC) analysis (Supplementary Materials and Methods). “All proteins” represent the percentage of all proteins excluding known DNA‐ and RNA‐binding proteins (Supplementary Materials and Methods). “DNA binding” proteins show the percentage of known poly‐nucleotide directly binding proteins (Supplementary Materials and Methods). Black bars indicate the standard deviation after considering replicates from three different experiments. Maria Lluch‐Senar et al. Mol Syst Biol 2015;11:780 © as stated in the article, figure or figure legend