Fibronectin Binding Is Required for Acquisition of Mesenchymal/Endothelial Differentiation Potential in Human Circulating Monocytes
Figure 3
Expression profiles of transcription factors in monocytes exposed to fibronectin or collagen (I) for 24 hours. (a) MACS-sorted CD14+ monocytes () were cultured with PBMC-conditioned medium on 24-well plastic plates coated with fibronectin (Sigma), collagen (I), or vehicle alone for 24 hours. Then, adherent cells were recovered and subjected to reverse transcription and semi-quantitative PCR for evaluation of mRNA expression. Positive controls for the specific amplification included cultured skin fibroblasts for Egr-1, HL-60 for , c-Myb, and AML-1, and THP-1 for IRF-7 and Foxp1. The results shown are representative of 5 independent experiments. (b) Quantitative Taqman real-time PCR examining gene expression levels of Egr-1 and AML-1 in adherent cells obtained from cultures of CD14+ monocytes with PBMC-conditioned medium for 24 hours on fibronectin (BD Biosciences) or collagen (I). Results obtained from 10 independent healthy donors are shown. Differences in expression levels are tested by paired t-test.
