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Fig. 4 | Arthritis Research & Therapy

Fig. 4

From: Nintedanib downregulates the transition of cultured systemic sclerosis fibrocytes into myofibroblasts and their pro-fibrotic activity

Fig. 4

Gene and protein expression of myofibroblast phenotype markers and ECM macromolecules in Scl70+ILD+ fibrocytes treated with nintedanib. A Quantitative real time polymerase chain reaction (qRT-PCR) of αSMA, S100A4, COL1, and FN in cultures of fibrocytes isolated from patients with anti-Scl70 positivity and ILD (Scl70+ILD+) maintained in normal growth medium without any treatment and treated with nintedanib at the concentrations of 0.1 μM and 1 μM for 3 h. The qRT-PCR is performed on eight independent in vitro experiments derived from fibrocytes obtained from each enrolled Scl70+ILD+ patient. Gene expression corresponds to the expression level (fold-increase) of the target gene of nintedanib-treated SSc fibrocytes compared with that of untreated cells, taken as the unit value by definition [18]. Data are expressed as median with range. B Western blotting and related densitometric analysis of protein synthesis of αSMA, S100A4, COL1, FN, and GAPDH in cultured Scl70+ILD+ fibrocytes maintained in normal growth medium without any treatment and treated with nintedanib at the concentrations of 0.1 μM and 1 μM for 24 h. Western blotting is performed on eight independent in vitro experiments. For each experimental condition, the value for the synthesis of αSMA, S100A4, COL1, and FN is normalized to that of the corresponding GAPDH. The resulting value of each treatment is compared with that of the related untreated cells (taken as unit value)

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