Fig 1: Effects of Tax on tumor growth of gastric cancer in vivo. (A and B) BALB/c null nude mice were administered AGS/NCI-N87 cell suspension injection to establish a gastric cancer animal model, and were treated with Tax. After sacrifice, immunohistochemical analysis was conducted to investigate the expression level of Ki67 of tumor tissues from mice injected with AGS or NCI-N87 cells. (C and D) The protein expression levels of Ki67, PCNA, MMP2, MMP9, E-cadherin, N-cadherin, ZO-1 and Snail were assessed using western blotting. (E and F) The protein expression levels of AhR and CYP1A1 were assessed using western blotting. ***P<0.001 vs. the control. Tax, taxifolin; PCNA, proliferating cell nuclear antigen; MMP, matrix metalloproteinase; ZO-1, Zonula occludens-1; AhR, aryl hydrocarbon receptor; CYP1A1, cytochrome P450 1A1.
Fig 2: Effect of Tax on AhR/CYP1A1 signaling in gastric cancer cells. (A) In AGS cells, the protein expression levels of AhR and CYP1A1 of each group were detected by western blotting. (B) In NCI-N87 cells, the protein expression levels of AhR and CYP1A1 of each group were detected by western blotting. **P<0.01 and ***P<0.001 vs. the control. Tax, taxifolin; AhR, aryl hydrocarbon receptor; CYP1A1, cytochrome P450 1A1.
Fig 3: Effects of AhR agonist, SB203580, on cell viability and proliferation in Tax-treated gastric cancer cells. (A and B) AGS and NCI-N87 cells were treated with Tax or co-treated with Tax and SB203580, an agonist of AhR. Then, the protein expression levels of AhR and CYP1A1 were assessed using western blotting and quantified. (C and D) The cell viability was assessed using a Cell Counting Kit-8 assay. (E) Cell colony formation assays were performed. ***P<0.001 vs. the control; #P<0.05, ##P<0.01 and ###P<0.001 vs. Tax. Tax, taxifolin; AhR, aryl hydrocarbon receptor; CYP1A1, cytochrome P450 1A1.
Fig 4: Effects of AhR agonist, SB203580, on cell migration and invasion in Tax-treated gastric cancer cells. (A) Wound-healing and Transwell assays were conducted to observe cell migration and invasion abilities, respectively. (B) The migration rate of each group was quantified. (C) The invasion rate of each group was quantified. (D and E) The protein expression of MMP2, MMP9, E-cadherin, ZO-1, N-cadherin and Snail was assessed by western blotting. ***P<0.001 vs. the control; ##P<0.01 and ###P<0.001 vs. Tax. Tax, taxifolin; AhR, aryl hydrocarbon receptor; CYP1A1, cytochrome P450 1A1; MMP, matrix metalloproteinase; ZO-1, Zonula occludens-1.
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