Fig 2. LPLI induces ROS-mediated RelA activation and BECN1 expression in human oral cancer cells.

(A) OECM-1 (white) and Ca9-22 (black) cells were pretreated with (+) or without (-) 10 mM NAC prior to irradiation with LPLI (810 nm, 60 J/cm²). The ROS production in LPLI-treated cells was determined using an ROS assay kit and 100 μM H₂O₂ as a positive control. (B) OECM-1 (white) and Ca9-22 (black) cells transfected with the NF-κB-responsive luciferase vector were pretreated with or without 10 mM NAC for 1 h then irradiated with LPLI (810 nm, 60 J/cm²). The treated cells were recovered for 6 h and 200 μM D-luciferin was added to monitor luciferase activity. (C) OECM-1 or Ca9-22 cells pretreated with (+) or without (-) 10 mM NAC were irradiated with LPLI (810 nm, 60 J/cm²) then recovered for 24 h. The recovered cells were harvested for immunoblotting to determine the protein levels of phosphorylated RelA, BECN1, and MAP1LC3-II. Protein levels for (D) phosphorylated RelA and (E) BECN1, and (F) the MAP1LC3-II/I ratio were quantified using ACTB as a normalization control. The data are expressed as the mean ± SEM from three independent experiments.