Figure 4. EBV induces myofibroblasts activation markers in infected-SSc-fibroblasts.
(a-b) Double IF of adherent fibroblasts stained with EBV-early/lytic-BFRF1 and αSMA-antigen (red) as indicated. Diaminidino-2-phenylindole (DAPI) was used as counterstaining for the nuclei (scale bar 50 μm). (c) Western-blot was performed to determine pSMAD2 and type I collagen secretion in cell-lysates and in the media of indicated fibroblasts cultures at 4-week-post-infection (PI). Total protein loading was determined by Ponceau-S staining. (d) Representative PCR products of EBV-DNA in 2 SSc-fibroblast-cell-lines infected with Monocytes (MO)-EBV-p2089 and dendritic-cells (DCs)-EBV-p2089 after 4 week PI; DNA from B95-8-EBV-positive cells were used as positive control. (e) mRNA expression of indicated genes in EBV-p2089-infected and in mock-infected-fibroblasts 4-week PI, evaluated by q-PCR. Bars represent mean ± S.E.M. from 5 separate SSc-fibroblast-lines. p-values calculated using two-tailed T-test