Fig. 2

Gene and protein expression of myofibroblast phenotype markers and ECM macromolecules in Scl70⁺ILD⁺ and SSc70⁻ILD⁻ fibrocytes. A Quantitative real time polymerase chain reaction (qRT-PCR) of αSMA, S100A4, COL1, and FN in cultures of fibrocytes isolated from nine SSc patients with anti-Scl70 antibody positivity and interstitial lung disease (Scl70⁺ILD⁺), five patients negative for anti-Scl70 without ILD (Scl70⁻ILD⁻), and five healthy subjects (HSs) maintained in normal growth medium without any treatment. Gene expression corresponds to the expression level (fold-increase) of the target gene of Scl70⁺ILD⁺and Scl70⁻ILD⁻ fibrocytes compared with that of HS fibrocytes taken as the unit value by definition [18]. Data are expressed as median with range. B Evaluation by Western blotting and related densitometric analysis of αSMA, S100A4 COL1, FN, and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) in cultures of fibrocytes isolated from nine Scl70⁺ILD⁺, five patients negative for anti-Scl70 without ILD (Scl70⁻ILD⁻), and five healthy subjects (HSs) maintained in normal growth medium without any treatment. For each sample, the values of αSMA, S100A4, COL1, and FN are normalized to that of the corresponding GAPDH. The resulting value of each sample of SSc fibrocytes is compared to that isolated from HS fibrocytes (taken as the unit value). Data are reported as median with range