Figure 4.

The NF-κB regulation by miRNA-dependent RSK2 suppression: (A) NF-κB promoter measurement using luciferase reporter assay. (B) Restored NF-κB activity by RSK2 overexpression in miR-29/-140-5p expressing A549. The cells were co-transfected by miR-29a/-140-5p and pcDNA-RPS6KA3 plasmid for 48h. Subsequently the promoter activity was measured by NF-κB promoter luciferase assay. Error bars indicate mean ±SD (n = 3) and the p-value was obtained by fisher exact t-test (*p < 0.05, **p < 0.01). (C) The phosphorylation of P65 subunit of NF-κB protein was analyzed by Western blot analysis. (D) Subcellular fractionation assay indicating nuclear localization of p65 depending on RSK2. Enhanced NF-κB activity by miR-29/-140 suppression. The H460 revertant was co-transfected by anti-miR-29/-140 and siRPS6KA3 siRNAs for 48h, and the promoter activities were addressed by (E) luciferase assay and (F) Western blot analysis. P-values were calculated by paired student t-test (*p < 0.05, **p < 0.005, ***p < 0.0005).