A Profibrotic Phenotype in Naïve and in Fibrotic Lung Myofibroblasts Is Governed by Modulations in Thy-1 Expression and Activation

Figure 3

Inverse correlation between Thy1 expression and fibroblast proliferation. (a) 24 h following transfection, cell mass was measured by colorimetric assay using methylene blue staining in Mlg cells transfected with Thy1 expression vector and in control plasmid-containing Thy1 cDNA in antisense orientation. Results are presented as fold change from day 0 ( in each group) (). (b) BrdU uptake in Mlg-transfected cells with Thy1 expression vector or control plasmid-containing Thy1 cDNA in antisense orientation following treatment with 20 μM BrdU for 2 h. BrdU uptake was assessed by anti-BrdU-FITC-conjugated mAb staining versus PI-total DNA staining followed by flow cytometry analysis. (c) In primary lung myofibroblasts transfected with nontargeting siRNA (Ctl) or Thy1 siRNA, cell mass was measured by colorimetric assay after methylene blue staining, 24 h posttransfection. Results are presented as fold change from day 0. In each group, , . (d) BrdU uptake in primary lung myofibroblasts transfected with nontargeting siRNA (Ctl) or Thy1 siRNA, following transfection and treatment with 20 μM BrdU for 6 h. BrdU uptake was assessed by flow cytometry analysis, following anti-BrdU-FITC-conjugated mAb staining versus PI-total DNA staining.

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