The DNA hypomethylating agent 5-aza-2′-deoxycytidine restores mRNA expression of Thy-1 that is accompanied by a decrease in α-SMA expression. Normoxic and hypoxic fibroblasts were treated with 1 μM 5-aza-2′-deoxycytidine (5-aza2dC) and quantitative real-time PCR was used to measure Thy-1 mRNA expression. A. 5-aza2dC inhibited hypoxia induced suppression of Thy-1 whereas in normoxia Thy-1 expression remained unchanged with 5-aza2dC treatment. B. Restoration of Thy-1 in 5-aza2dC treated hypoxic cells was accompanied by a reduction in α-SMA mRNA. C. Immunocytochemistry for α-SMA (red) demonstrated that restoration of Thy-1 using 5-aza2dC treatment decreased expression of α-SMA in hypoxic cells. Cells were counterstained with nuclear specific DAPI (blue). Results are mean +/- SD for n = 3 experiments and students t test was used for statistical analysis.