FIG 11.

Stable transduction of HEK293 or HeLa cells with lentiviral shRNA vectors targeted to eGFP. (a and b) HEK293 or HeLa cells were stably transduced with lentiviral shRNA vectors targeting eGFP at an MOI of 2, after which independent cellular clones were isolated. Untreated HEK293 or HeLa cells were used as controls. The indicated cells were infected with lentiviruses expressing eGFP at an MOI of 2. The protein level of eGFP was measured by Western blotting 48 h after infection. Actin was used as the protein-loading control. (c) Relative amounts of miR-216b-5p and miR-30b-5p were examined in the stably transduced HEK293 pool and cell clones (HEK293-8 and HEK293-42) by real-time PCR. Untreated HEK293 cells were used as controls. (b) Relative amounts of miR-216b-5p and miR-30b-5p were examined in the stably transduced HeLa pool and cell clones (HeLa-3 and HeLa-4) by real-time PCR. Untreated HeLa cells were used as controls. Because miR-216b-5p was not detected in the untreated HeLa cells, the relative amount of miR-216b-5p in the transduced cells is presented as 2^−ΔΔCT. (e and f) The protein levels of JUN in stably transduced pools and clones were measured by Western blotting. Actin was used as a loading control. *, P < 0.05 (considered significant) (t test; n = 3).