FIG. 5.

(A) Oligonucleotide uptake by primary cytotrophoblasts. After 5 h of culture, cytotrophoblasts were incubated with FITC-labeled scrambled oligonucleotide for 1, 2, 24, and 48 h. For each time point, cells were washed three times in PBS, fixed, and analyzed by fluorescence microscopy. Nuclei were stained in blue by DAPI. Intensely fluorescent green cells have internalized the scrambled antisense oligonucleotide (FITC). (B) Evaluation of Env-W antisense oligonucleotide. After 5 h of culture, cytotrophoblasts were incubated with three specific env antisense oligonucleotides (Env-W 744-766, Env-W 750-770, and Env-W 758-777) or a random oligonucleotide (scrambled) or without oligonucleotide (control). After 48 h of culture, proteins were extracted and Env glycoprotein expression was analyzed by Western blotting using an anti-SU polyclonal antibody and standardized using an antiactin monoclonal antibody. The most efficient reduction of Env expression was obtained with the Env-W 758-777 antisense oligonucleotide.