Fig. 4.

Tp0768 regulated the production of proinflammatory cytokines in cells through the ROS/NF-κB pathway. a NF-κB (p65) was detected using western blotting, and its gray value was analyzed using ImageJ software. b Nuclear translocation of NF-κB (p65) was assessed using immunofluorescence staining and confocal microscopy. DIPA was used for nuclear staining at a scale of 5 μm. c NF-κB (p65) levels were detected using western blotting in the absence or presence of NAC. The relative mRNA levels of IL-1β, IL-6, and IL-8 in THP-1–differentiated macrophages (d) and Raw264.7 cells (e) were detected using RT-PCR in the absence or presence of PDTC, respectively. Values are expressed as fold changes relative to GAPDH normalized mRNA levels. All data are presented as mean ± SD of at least three independent experiments. *p < 0.05, **p < 0.01, and ***p < 0.001 indicate a significant difference from the control group