Figure 4.

TPL enhanced GEM-mediated apoptosis of PANC-1 cells by inhibiting TLR4/NF-κB signaling. (A) PANC-1 cells were incubated with 0.01, 0.1, 1, or 10 μg/mL GEM and 40 ng/mL TPL for 48 h or pretreated with 1 μg/mL of LPS for 4 h, followed by incubation with TPL or GEM for 48 h. Cell proliferation was assessed by MTT assay. (B) PANC-1 cells were incubated with 1 μg/mL GEM and 40 ng/mL TPL for 24 h or pretreated with 1 μg/mL of LPS for 4 h, followed by incubation with TPL or GEM for 24 h. Cell apoptosis was determined by Annexin V staining followed by flow cytometry. (C) Representative histograms for the flow cyctometric data in (B). (D) Cells were treated as in (B), and the relative activity of NF-κB was detected using a double luciferase reporter gene. (E) Cells were treated as (B), and total proteins were extracted for western blotting of the indicated proteins with β-actin as the loading control. *P < 0.05; **P < 0.01.