Figure 1

Generation and characterization of a one-cycle measles virus (MV) vector expressing OCT4. (A) Top: MV genome. Middle: Genome of MV expressing green fluorescent protein (GFP) but no H protein. Bottom: Genome of MV expressing OCT4 and GFP but no H protein. The MV antigenome (plus strand) is represented with its 5′ end on the left; the six genes are indicated by capital letters. (B) Flow cytometry analysis of H expression on 293-3-46 (left) and Vero (right) cells after LV-H transduction. Top graphs, mock-transduced cells. Bottom graphs, LV-H transduced cells. (C) Immunoblot analysis of OCT4 expression. BJ and Vero cells were infected with the indicated vector and after 36-hour cell extract were analyzed by SDS-PAGE. OCT4 antibody against OCT4 protein was used. Uninfected BJ and Vero cells (Mock) were used as controls. β-actin antibody was used as control of protein load. (D) Immunofluorescence analysis of OCT4 expression. Vero and BJ cells were infected with the indicated vector for 36 hours and analyzed by immunostaining and confocal microscopy. The cells were fixed, permeabilized, and stained with antibody to OCT4 (red). Nuclei were counterstained by 4′,6-diamidino-2-phenylindole (DAPI) (blue). GFP (green) was expressed during infection. Scale bars: 100 μm. (E) Titers of cell-associated and released virus produced upon infection of Vero cells with MV(OCT4) or MV, determined at 24 hours (gray columns) or 48 hours (black columns) post-infection. (F) Titers of cell-associated and released virus produced upon infection of Vero-H2 cells with MV(OCT4) or MV, determined at 24 hours (gray columns) or 48 hours (black columns) post-infection.