MOESM3 of Pyruvate kinase M2 and the mitochondrial ATPase Inhibitory Factor 1 provide novel biomarkers of dermatomyositis: a metabolic link to oncogenesis

Additional file 3: Figure S2. Overexpression of PKM2 in DM. a Tissue extracts (30 µg) derived from two randomly selected muscle biopsies of control donors (CTR), polymyositis (PM), dermatomyositis (DM) and sporadic inclusion body myositis (IBM) and of the HCT116 cell line were fractionated on SDS-PAGE and blotted against anti-PKM2, anti-GAPDH and anti-LDH-A. Electrophoretic migration of the protein is indicated to the left of each blot. b Histograms show the expression of the proteins (a.u.) when compared to the expression in HCT116 cells

MOESM1 of Pyruvate kinase M2 and the mitochondrial ATPase Inhibitory Factor 1 provide novel biomarkers of dermatomyositis: a metabolic link to oncogenesis

Additional file 1: Figure S1. Linear correlation between the fluorescence intensity and the content of native proteins. HCT116 cell line extracts (0–1 μg/μl) were spotted in the arrays (see Fig. 2a). Significant linear correlations were obtained between the fluorescence intensity (arbitrary units, a.u.) of the spots and the amount of the protein interrogated in the arrays. Protein concentrations in the biopsies were calculated by interpolation in the respective linear plots

MOESM4 of Pyruvate kinase M2 and the mitochondrial ATPase Inhibitory Factor 1 provide novel biomarkers of dermatomyositis: a metabolic link to oncogenesis

Additional file 4: Figure S3. Plasma levels of PKM2 by ELISA and RPPA. a Determination of PKM2 by ELISA in plasma samples of DM, sIBM and CRL patients. b Upper panel, scheme of printing of the plasma samples from CRL, sIBM, DM and PM patients. One nl of 1:20 diluted samples were spotted in quadruplicate. Black boxed: negative controls of BSA; Magenta boxed: standard curves of HCT116 cells; Brown boxed: positive controls of murine IgGs; Orange boxed: standard curve of PKM2 recombinant protein; Green boxed: samples from control donors (CTR); Blue boxed: samples from sIBM patients; Red boxed: samples from DM patients; Yellow boxed: sample from PM patients. Lower panel, parallel array processed with goat anti-mouse IgGs CF647 as negative control. Please note the lack of cross-reactivity against the human plasma IgGs. c The histogram shows the plasma levels of PKM2 quantified by RPPA assay. The results shown are mean ± S.E.M

Additional file 1: Figure S1. of RIG-I expression in perifascicular myofibers is a reliable biomarker of dermatomyositis

Controls used in the study to assess the specificity of the RIG-I antibody. RIG-I antibody was incubated in non-transfected HEK293 cells (left) and in HEK293 cells transfected with the constitutive expression vector pCMV-RIG-I (right). Positivity was only observed in those cells transfected with pCMV-RIG-I. Original magnification × 400. (TIF 3652 kb