Fig. 4.

Effects of morpholinos on in vitro ovine trophectoderm growth. Mononuclear trophectoderm cells were isolated from day 15 conceptuses. (A) Morpholino delivery. Rhodamine-labeled MAO-std was complexed with Endo-Porter aqueous delivery reagent and added to cells in culture. Fluorescence microscopy was used to visualize the labeled MAO in cells. [Width of each field of view is 870 μm (Left) and 90 μm (Right).] (B) Effect of morpholinos on enJSRV Env and Gag protein in cultured trophectoderm cells. Cells were grown on glass slides and mock-treated or treated with MAO-std, MAO-5mis, or MAO-env for 48 h. Immunofluorescence analysis determined that synthesis of enJSRV Env, but not Gag, protein was inhibited in cells treated with MAO-env but not the other morpholinos. Results are representative of three experiments. (Width of each field of view is 140 μm.) (C) Effect of morpholinos on trophectoderm cell growth. Cells were grown in culture dishes until 30% confluency and mock-treated (no morpholino) or treated with MAO-std, MAO-5mis, or MAO-env for 48 h. Cell number was reduced (P < 0.05) by 33% in cultures treated with the MAO-env relative to control morpholinos. Results are from three independent experiments, and data are expressed as the percentage of cell number in mock-treated cultures.