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. 2021 Feb 24;10:623048. doi: 10.3389/fonc.2020.623048

Figure 5.

Figure 5

Tumor necrosis factor-induced protein-8 (TIPE)-mediated activation of the PI3K/Akt pathway is involved in the modulation of DcR3 levels in HCT116 cells. (A) HCT116 cells were transfected with TIPE or an empty vector (control) for 24 h with lipopolysaccharide (LPS) stimulation for the indicated time (0, 15, 30, and 60 min). Cells were harvested, and whole-cell extracts were prepared for Western blot analysis of the indicated proteins. The blots shown are representative of those obtained in three separate experiments. (B) HCT116 cells were cultured in 6-well plates and transfected with TIPE or an empty vector (control) for 24 h, then qRT-PCR assays were performed to measure the relative DcR3 mRNA expression of HCT116 cells after treatment with LY294002 (50 μM), NSC23766, U0126, and BAY 11-7082. Expression was normalized to that in the control cells. (C) The culture medium was collected, and DcR3 protein levels were measured by ELISA. (D) HCT116 cells were transfected with tumor necrosis factor-induced protein-8 (TIPE) after treatment with lipopolysaccharide (LPS) for the indicated time (0, 30, and 60 min), and Western blot analysis of whole-cell lysates was performed to examine the indicated proteins in HCT116 cells after treatment with or without LY294002 (50 μM). ns, not significant, *p < 0.05, ***p < 0.001.