Replication in Cells of Hematopoietic Origin Is Necessary for Dengue Virus Dissemination
Figure 2
Generation and characterization of miR-142-targeted DENV-2.
(A) Cloning strategy for the insertion of miR-142 target sites into the 3′untranslated region (UTR) of a T7-driven DENV-2 cDNA clone. A 157 nucleotide (nt) insert containing four tandem target sites were cloned into the variable region of DENV-2 (142t virus). A control (ctrl) virus containing four reverse sites is also depicted. (B) Northern blot for miR-142 expression in Aedes albopictus mosquito (C6/36) cells, mammalian baby hamster kidney cells (BHKs), murine bone-marrow-derived macrophages (BMMs), and a human B cell line (Raji). (C) Quantitative RT-PCR and western blot analysis on C6/36 cells infected with wt, ctrl, and 142t viruses at the indicated time points. Wild type (wt) virus refers to a clone encompassing no modifications. (D) Same as described in (C) for BHK cells.