Figure 3

Effects of various STAT3 pathway inhibitors on everolimus-mediated apoptotic effects and cell growth inhibition in HaCaT cells. (A) HaCaT cells were incubated in medium containing everolimus at the indicated concentrations for 48 h after pretreatment with 10 μM stattic or DMSO for 20 min. Subsequently, apoptotic cells were detected using FITC-labeled Annexin V/PI staining on an IN Cell Analyzer 2000 for Imaging cytometric analysis. (B) Effects of JAK/STAT pathway inhibitors and IL-6 on the cell growth inhibition induced by everolimus. HaCaT cells were incubated in medium containing 30 μM everolimus for 48 h after pretreatment with 10 μM stattic for 20 min or coincubation with everolimus and 25 μM Z3 (a selective inhibitor of JAK2), 20 μM STA-21, 100 ng/mL IL-6, or DMSO (solvent of these inhibitors). Cell viability was determined by WST-8 colorimetric assay.