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Fig. 3 | Arthritis Research & Therapy

Fig. 3

From: Blocking CD248 molecules in perivascular stromal cells of patients with systemic sclerosis strongly inhibits their differentiation toward myofibroblasts and proliferation: a new potential target for antifibrotic therapy

Fig. 3

Transforming growth factor (TGF)-β and platelet-derived growth factor (PDGF)-BB effects on CD248, α-smooth muscle actin (α-SMA), and Ki-67 expression in systemic sclerosis (SSc) mesenchymal stem cells (MSCs). a qRT-PCR of CD248 messenger RNA (mRNA) levels in ten SSc-MSC (five early-onset subset [EOS] and five long-standing subset [LSS]) and ten healthy control subject (HC) MSC samples. In SSc-MSCs, CD248 mRNA expression levels are always significantly higher than in HC-MSCs. b qRT-PCR of α-SMA mRNA levels in ten SSc-MSCs (five EOS and five LSS) and ten HC-MSCs. In SSc-MSCs, the α-SMA mRNA levels are always significantly higher than in HC-MSCs. In both SSc- and HC-MSCs, TGF-β treatment induces a significant increase of α-SMA mRNA expression compared with untreated (UT) cells. On the contrary, PDGF-BB treatment induces a significant decrease of α-SMA compared with UT cells in both HC- and SSc-MSCs. c qRT-PCR of Ki-67 mRNA levels in ten SSc-MSCs (five EOS and five LSS) and ten HC-MSCs. In both SSc- and HC-MSCs, TGF-β treatment induces a significant decrease of Ki-67; on the contrary, PDGF-BB induces a significant increase of Ki-67 when compared with UT cells in both SSc- and HC-MSCs. The TGF-β isoform used is TGF-β1. Any single dot in the figure represents the median of triplicate experiments for each patient ** p = 0.0002, *** p = 0.0001. d Western blot analyses performed in four SSc-MSCs (two EOS and two LSS) and four HC SSc-MSCs confirmed the results observed by qRT-PCR analyses. Pictures are representative of all experiments. e and f Densitometric analysis of (e) CD248 protein bands and (f) α-SMA protein bands. The values were expressed as protein relative quantification/β-actin relative quantification. * p = 0.02

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