Fig. 3
GTS-21 suppressed TNF-α production in macrophages after LPS-stimulation in vitro. (A) Experimental design: 1.0 × 105 RAW 264.7 cells were seeded in 24-well plates for 24 hours. TNF-α was induced by LPS (100 ng/mL). Four and 6 hours later, GTS-21 (50 μM) was administered, and eight hours later, the supernatant was collected. (C) Experimental design: 1.0 × 105 U937 cells were seeded in 24-well plates for 24 hours. TNF-α was induced by LPS (1 μg/mL). Four and 6 hours later, GTS-21 (50 μM) was administered, and 8 hours later, supernatant was collected. (B, D) TNF-α concentrations were analyzed by ELISA. The decrease in TNF-α was observed with GTS-21 administration in both RAW 264.7 (B) (vehicle-vehicle and LPS-vehicle; P < 0.0001, LPS-vehicle and LPS-GTS-21; P < 0.0001) and U937 cells (D) (vehicle-vehicle and LPS-vehicle; P = 0.0003, LPS-vehicle and LPS-GTS-21; P = 0.0093). Relative TNF-α levels were calculated based on the average value of TNF-α increased by LPS stimulation (D). Data are shown as mean ± SD. Statistical significance was assessed using two-way ANOVA analysis with Tukey’s multiple comparison. **P < 0.01, ***P < 0.001,****P < 0.0001.
