Research Paper Volume 12, Issue 22 pp 23114—23128

Figure 3. Intracellular S100A9 regulates phosphorylation of AKT1^Thr308 and induces EMT in PA. (A) The mRNA levels of S100A9 in control group (group 1), NC1 group (group 2), LV-S100A9 group (group 3) were detected by Real-Time PCR(n=9, P^***<0.001 versus group 1). (B) Western blot was used to observe S100A9 protein levels in groups 1, 2, and 3(n=5, P^***<0.001 versus group 1). (C) The expression of phospho-AKT1^Thr308, Vimentin, ICAM-1 and E-cadherin was explored using western blot analysis in groups 1, 2, 3 and a LV-S100A9 + addition of A-674563 group (group 4)(n=5, P^***<0.001 vs. group 1. P^##<0.01; P^###<0.001 vs. group 4). (D, E) Real-time PCR and western blot analysis were used to investigate S100A9 mRNA and protein expression in the control group (group a), NC2 group (group b) and S100A9-shRNA-LV group (group c), respectively(n=5, P^***<0.001, compared with group a). (F) Western blot demonstrated that downregulation of S100A9 reduced phospho-AKT1^Thr308, Vimentin, ICAM-1, coupled with increase in E-cadherin. The S100A9-shRNA-LV + SC79(AKT1 agonist) group (group d) displayed that the reduced phospho-AKT1^Thr308, Vimentin, ICAM-1 and elevated E-cadherin were reversed partially by SC97(n=5. P***<0.001, compared with group a. P##<0.01; P###<0.001, compared with group d).