Fig. 5.

Effect of PTEN-knockdown (KD) on Akt phosphorylation and cytokine production. BEAS-2B cells were incubated with random oligonucleotide (RO; 100 nM) or short interference (si) RNA against PTEN (100 nM) for 48 h. The levels of PTEN were determined by RT-qPCR (A) and Western blot analysis (B). The ratio of p-Akt/total-Akt as well as PTEN/β-actin were also determined (n = 3). Effects of PTEN knockdown on cytokine production determined by cytokine array (C–E) or by ELISA for IL-6 (F), CXCL8 (G), CCL5 (H) and CCL2 (I). J: effect of stimulation with IL-1β (0, 0.01, 0.1, and 1 ng/ml) on CXCL8 release for 24 h in PTEN-KD cells. An impact of PTEN KD on gene expression IL-6 (K), CXCL8 (L), matrix metalloproteinase-9 (MMP-9; M), tranforming growth factor-β (TGF-β; N), MUC5AC (O) and MUC5B (P), all which were corrected with the gene expression of a housekeeping gene GNB2L1 (n = 3). All values are mean values ± SE of 3-6 separate experiments. *P < 0.05, **P < 0.01, compared with the values of RO-treated group; ^††P < 0.01, compared with the values of nontreatment RO group; ^‡P < 0.05, ^‡‡P < 0.01, compared with the value of nontreatment PTEN-KD group. ^§§P < 0.01, between the RO group and PTEN-KD group.