Figure 4. Transfection with low-dose 3p-RNA induces activation of NF-κB, MAPK and Akt.

(A) CAL27 cells (2×10⁶ per well) were cultured in 6-well plates and transfected with indicated doses of 3p-RNA or BE-RNA for 16 h. Cell lysates were collected and analyzed by Western blot using the indicated antibodies. (B) Nuclear extracts prepared from CAL27 cells transfected as in (A) were subjected to Western blot analysis with anti-NF-κB p65, IRF3 and AP-1 antibodies. (C) CAL27 cells transfected as in (A) were treated with indicated inhibitors for 12 h and then collected for the MTT assay. (D) CAL27 cells (2×10⁶ per well) were cultured in 6-well plates and transfected with indicated doses of 3p-RNA or BE-RNA for 16 h. Cell lysates were collected and analyzed by Western blot using the indicated antibodies. Results are presented as the absorption ratio. Similar results were obtained in multiple repetitions (at least three) of these experiments.