TRAF6 inhibition suppresses the IL-33/ST2-triggered vasculogenic mimicry (VM) and epithelial–mesenchymal transition (EMT) in vitro (IMAGE)

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TRAF6 inhibition suppresses the IL-33/ST2-triggered vasculogenic mimicry (VM) and epithelial–mesenchymal transition (EMT) in vitro

Caption

(A) Western blotting analysis of EMT-related markers in oral squamous cell carcinoma (OSCC) cells treated with IL-33. (B) mRNA and protein expression levels of EMT-related markers were detected via quantitative PCR (left) and Western blotting analysis (right) in IL1RL1-knockdown and control OSCC cells treated with or without IL-33. n = 3 biological replicates. (C) Western blotting analysis of OSCC cells treated with IL-33 for the indicated times. (D) Western blotting analysis of OSCC cells treated with IL-33 alone or combined with C25-140. (E) Western blotting analysis of TRAF6-overexpressing SCC1 cells and corresponding control cells treated with IL-33 alone or combined with C25-140. (F) Immunofluorescence analysis of p65 nuclear translocation in TRAF6-overexpressing SCC1 cells and corresponding control cells treated with IL-33 alone or combined with C25-140. The quantification of immunofluorescence analysis (right). n = 3 biological replicates. (G) Wound healing assay for SCC1 cells treated with IL-33 alone or combined with C25-140. The quantitative statistics of the migration rate (right). n = 3 biological replicates. (H) Tube formation assay of TRAF6-overexpressing SCC1 cells and corresponding control cells treated with IL-33 alone or combined with C25-140. The quantitative plot of the VM-related parameters (right). n = 3 biological replicates. (I) mRNA and protein levels of VM-, EMT-related markers, and HIF-1α in SCC1 cells treated with IL-33 alone or combined with C25-140. n = 3 biological replicates. Data were presented as the mean of three independent experiments ± standard deviation. In (B, F, G, H, I), one-way ANOVA with multiple comparison post hoc analysis was used to determine P-values. ns, no significance; ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001, and ∗∗∗∗P < 0.0001. Scale bars, 50 μm.

Credit

Wendong Wan, Liwei Ma, Wenjin Wang, Jianjun Wu, Chengxin Shi, Xuancheng Hui, Muhammad Tufail, Cheng Guo, Canhua Jiang, Lunquan Sun, Ning Li, Xing Gao

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