Effects of CQ on the levels of LC3B, anti-apoptotic proteins, and DRs, of two human pancreatic cancer cell lines.

<p>(A, B) Two cancer cell lines were cultured with CQ (200 nM for MiaPaCa-2 and 700 nM for Panc-1) and/or TRAIL (10 ng/mL for MiaPaCa-2 cells, 100 ng/mL for Panc-1 cells) for 24 h, lysed, and the expression levels of LC3B, p62, and Beclin-1 were measured. β-Actin and α-tubulin served as controls. (C) Two cancer lines were cultured in the presence of CQ (200 nM for MiaPaCa-2 and 700 nM for Panc-1). After 48 h, DR5 and DR4 expression levels were examined by flow cytometry. The lines exhibited binding of mAbs specific for either DR5 or DR4, followed by binding of an FITC-conjugated secondary antibody. The solid gray indicates binding of an isotype-matched control mouse IgG, followed by binding of an FITC-conjugated secondary antibody. (D) Two cancer cell lines were cultured with CQ (200 nM for MiaPaCa-2 and 700 nM for Panc-1 cells) for 48 h, and the expression levels of cFLIP, Bcl-xL, and XIAP were measured. β-Actin served as a control. The number represents the band density of each protein, which was normalized to the β-actin loading control. Similar results were obtained in two independent experiments.