Fig. 4: Regulation of TF mRNA 3′-UTR by vimentin. | Oncogene

Fig. 4: Regulation of TF mRNA 3′-UTR by vimentin.

From: Vimentin prevents a miR-dependent negative regulation of tissue factor mRNA during epithelial–mesenchymal transitions and facilitates early metastasis

Fig. 4

a RNA-immunoprecipitation assays. a MDA-MB-231 cell lysates were subjected to immunoprecipitation with a vimentin antibody (IP Vim). RNA was extracted from the immunoprecipitate and subjected to RT-qPCR for TF, 18S or TBP. b MDA-MB-231 cell lysates were subjected to immunoprecipitation with a vimentin (IP Vim) or a cytokeratin 8/18 antibody (IP CK8/18) and extracted RNAs were subjected to RT-qPCR for TF. Results from a representative experiment are shown. c Cell lysates from Ctrl or TGF-β1-treated A549 cells were subjected to immunoprecipitation with a vimentin antibody (IP Vim). RNA was extracted from the immunoprecipitate and subjected to RT-qPCR for TF. Immunoprecipitation conditions with control IgG were included in all experiments. b TF 3′-UTR-luciferase reporter assays. MDA-MB-231, EGF-treated MDA-MB-468 and TGF-β1-treated A549 cells were transfected with a reporter vector containing the firefly luciferase coding sequence inserted upstream the 3′-UTR of TF mRNA. The vector also expresses the renilla luciferase as an internal control. Results are expressed as the ratio firefly luciferase/renilla luciferase.

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