Figure 1

Cav-1 modulation in SSc-MSCs. (A) Immunofluorescence staining of Cav-1 in limited and diffused SSc-MSCs and HC-MSCs. The pictures showed an impairment of the Cav-1 trafficking in SSc cells. Pictures are representative of all experiments. Original magnification 20X. (B) Cav-1 mRNA levels of expression. After 15 min of VEGF treatment, the mRNA Cav-1 levels were markedly increased, in both HC- and SSc-MSCs, and the values were significantly higher in HC-MSCs vs. SSc-MSCs. Each experimental condition was performed in triplicate. All the results are expressed as Median (range) ***P <0.0001. (C) Western blot analysis of Cav-1 and Cav-1-phosphorilation (p-Cav-1) in HC- and SSc-MSCs, using neutral detergent. The western blot analysis confirmed the decrease of Cav-1 expression in SSc-MSCs and showed that VEGF induced a significant upregulation and a tyrosine phosphorylation of Cav-1, although the levels were significantly lower in SSc-MSCs when compared with HC cells. Each experimental condition was performed in triplicate. Blot was representative of all the experiments. The values were expressed as arbitrary unit of optical density (OD) (**P <0.001; ***P <0.0001). (D) Western blot analysis of Cav-1 and Cav-1-phosphorilation (p-Cav-1) in HC- and SSc-MSCs, using SDS. The results obtained by using a different buffer mirror those obtained by neutral detergent. The values were expressed as arbitrary unit of OD (**P <0.001; ***P <0.0001).