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Ce coverage ( ) 25 47 33dcSSc/SRC1100 1420 1636Calreticulin precursor (T) Pre-mRNA splicing factor SPF27 (N) Eukaryotic translation initiation factor 5A-1 (N) Eukaryotic translation initiation factor 5A-1 (T) Probable ATP-dependent RNA helicase DDX17 (N)48/63 26/25 17/16 17/17 72/76 39/39 37/35 -4.3/4.4 5.5/5.9 5.1/5.7 5.1/5.dcSSc/PAH dcSSc/ILD 589 11018.8/8.0 6.3/6.9 5.9/6.8/20 5/10 10/207 13235 5636 41Poly(rC)-binding protein 2 (N) [SwissProt: PCBP2_HUMAN] Serine/threonine protein phosphatase PP1-a catalytic subunit (N) DNA-directed RNA polymerases I, II and III, subunit RPABC1 (N) Cofilin 1 (T) Histone H2A type 1-J (T) Telomeric repeat binding factor 2-interacting protein 1 (N) Heterogeneous nuclear ribonucleoprotein A/B (N) Peroxiredoxin 2 (T) 78-kDa glucose-regulated protein precursor (T) [SwissProt: PP1A_HUMAN] [SwissProt: RPAB1_HUMAN] [SwissProt: COF1_HUMAN] [SwissProt: H2A1J_HUMAN] [SwissProt: TE2IP_HUMAN] [SwissProt: ROAA_HUMAN] [SwissProt: PRDX2_HUMAN] [SwissProt: GRP78_HUMAN]dcSSc*25/5.7/6.2/2163 lcSSc/DU lcSSc/PAH 231719/19 14/16 44/8.2/9.5 10.9/6.1 4.6/4.3/7 2/3 9/134 3772 2054 271119 2079 lcSSc/ILD 901 2063 lcSSc* 820 147836/38 22/23 72/76 70/70 55/57 37/44 22/8.2/6.5 5.7/6.0 5.1/5.4 6.2/6.9 5.6/6.4 4.5/4.6 5.3/5.3/5 5/7 13/29 3/14 3/7 3/7 1/55 143 711 89 112 8227 40 121 45 64 3915 26 28 29 16 29ATP-dependent DNA helicase [SwissProt: 2, subunit 1 (N) KU70_HUMAN] U4/U6 small nuclear ribonucleoprotein Prp31 (N) Calumenin precursor (T) Tumour protein D54 (T) [SwissProt: PRP31_HUMAN] [SwissProt: CALU_HUMAN] [SwissProt: TPD54_HUMAN]a ANA: antinuclear antibody; dcSSc: diffuse cutaneous systemic sclerosis; DU: digital ulcer; ILD: interstitial lung disease; lcSSc: limited cutaneous systemic sclerosis; MW: molecular weight (in kilodaltons); N: proteins recognised in HEp-2 cell-enriched nuclear protein extract; PAH: pulmonary arterial hypertension; SRC: scleroderma renal crisis; SSc: systemic sclerosis; T: proteins recognised in HEp-2 cell total protein extract; th/es: theoretical/estimated. bNumber of unique identified peptides in MS/MS and in MS+MS/MS searches. cWithout visceral involvement.pathogenesis of SSc, we thought it more appropriate to use these cells as sources of autoantigens because we were looking for additional targets to ANAs. Additional validation studies with sera from patients with other connective tissue diseases are necessary. In addition, 2DE and immunoblotting were not adapted to test alarge number of sera, and thus further experiments using ELISA with recombinant proteins are necessary, which will allow for validation of the target antigens and screening PubMed ID:https://www.ncbi.nlm.nih.gov/pubmed/16989806 of a large number of patients. However, our work has several additional limitations. Less than 1,000 protein spots were stained in theBussone et al. Arthritis Research Therapy 2011, 13:R74 http://arthritis-research.com/content/13/3/RPage 10 ofFigure 4 Signalling network of proteins identified as major targets of autoantibodies in patients with unidentified ANA. This schematic representation, created by using Pathway Studio software, shows the connectivity between TGF-b and HEp-2 cell proteins identified as major targets of autoantibodies in SSc patients with unidentified ANA. Protein Capecitabine entities belonging to different functional groups are represented as different shapes. ANA: antinuclear antibody; CALR: calreticulin; CFL1: cofilin 1; FUS: fused in sarcoma; HDAC2: histone deacetylase 2; HNRNPA1: heterogeneous nuclear ribonucleoprotein A1; HNRNPA2B1: heterogen.