Fig. 4

Piezo1 channel activation mediated the pro-inflammatory responses via an increase in the release of CXCL8 into the tumor microenvironment. (A) Dual luciferase reporter assay recorded the luciferase activity in the Huh7 cells transfected with GPL4-Basic and GPL4-CXCL8 plasmids. (B) Using the Huh7 cells transfected with GPL4-Basic and GPL4-CXCL8 plasmids, the luciferase activities were examined in the Huh7 cells pre-treated with 1 µM SCH772984 and 10 µM MK-2206 for 2 h, and then treated with 10 µM Yoda1 for 24 h thereafter. (C) Schematic diagram describing the experimental procedure of the PBMC migration assay in vitro. (D) PBMC migration assay in the presence of hrCXCL8 (10, 100 and 1000 ng/mL). (E) PBMC migration assay in the presence of conditioned medium cultured with or without Huh7 and HepG2 cells exposed to 10 µM Yoda1 for 24 h. (F) Tube formation assay using HUVECs cultured with 1% DMSO, or 10 µM Yoda1, or hrCXCL8, or conditioned medium derived from Huh7 treated with 10 µM Yoda1 for 8 h. (G) The mRNA levels of VEGF-A in Huh7 and HepG2 cells after treatment with 10 µM Yoda1 for 24 h. (H) ELISA assay analyzed the release of VEGF protein in the medium of Huh7 and HepG2 cells after treatment with 10 µM Yoda1 for 24 h. *P < 0.05, ***P < 0.001; #P < 0.05, ###P < 0.001