Figure 2.

IL-6 intracellular levels in PEA1 and PEA2 cells. Cells (1×10⁵/mL) were incubated in RPMI 1640 medium at 37°C in a 5% CO₂ atmosphere for 6 hrs. (A) Supernatants were collected and analysed for IL-6 content by ELISA. (B) Effect of AKT or JNK inhibitors on IL-6 release by PEA1 and PEA2 cells. Cells (1×10⁵/mL) were incubated in RPMI 1640 medium, in the presence or absence of 4 μM AKT inhibitor (AKT Inhibitor IV, Calbiochem) or 10 μM JNK inhibitor (SP600125, Sigma-Aldrich) at 37°C in a 5% CO₂ atmosphere for 6 hrs. Cell lysates were analysed by western blot with an anti-IL-6 (ab6672, Abcam), an anti-pAKT (Ser473) (#9271, Cell signaling), an anti-AKT (#9272, Cell signaling) or an anti-GAPDH (sc-32233, Santa Cruz Biotechnology) antibody. (C) Supernatants from (B) were collected and IL-6 content was analysed by ELISA. All data were obtained from triplicate samples and confirmed in three separate experiments.