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PCR Cloning and Subcloning> | Restriction enzyme analysis and direct DNA sequencing confirmed the correct insertion of short hairpin RNA (shRNA) sequences. ShRNA targeting Ptk-2 (shFAK, TRCN0000023485) or Itgb1 (shITGB1, TRCN0000066645) mRNA on a self-inactivating lentiviral vector (pLKO.1 and pLL3.7, respectively) with a CMV driven tGFP (pLKO.1) or eGFP (pLL3.7) reporter were acquired from Sigma and Genscript, respectively (Table S2). | Get A Quote |
Tissue inhibitor of metalloproteinases-1 (TIMP-1) exerts pleiotropic effects on cells including conferring metastatic properties to cancer cells. As for metastatic cells, recent paradigms of leukocyte migration attribute important roles to the amoeboid migration mode of dendritic cells (DCs) for rapid locomotion in tissues. However, the role of TIMP-1 in immune cell migration and in the context of infection has not been addressed. We report that, upon challenge with the obligate intracellular parasite Toxoplasma gondii, primary DCs secrete TIMP-1 with implications for their migratory properties. Using a short hairpin RNA (shRNA) gene silencing approach, we demonstrate that secreted TIMP-1 and its ligand CD63 ar... More