TLR4 binds with PIEZO1 to promote calcium influx in BON-1 cells. (IMAGE)
Caption
(A) The mRNA expression levels of TLR4, PIEZO1, and TRPA1 in BON-1 cells treated with different concentrations of LPS were determined by quantitative PCR analysis (n = 5 per group; one-way ANOVA). (B) Calcium images showing calcium influx changes in the BON-1 cells treated with PBS or LPS combined with or without the TLR4 inhibitor TAK-242. Scale bar = 50 μm (n = 3 per group). (C) The co-localization observation of TLR4 (green) and PIEZO1 (red) in the four different treatment groups of BON-1 cell by confocal microscopy. Scale bar = 100 μm (n = 10 per group). (D) TLR4 bound to PIEZO1 in the BON-1 cell treated with or without LPS as determined by co-immunoprecipitation using TLR4 or PIEZO1 antibodies. (E)The concentration of 5-HT in the supernatant of BON-1 cells treated with LPS or PBS, with or without treatment with TAK-242 (n = 10), was determined by ELISA. Data were expressed as mean ± standard error of the mean; ns, not significant; ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001, and ∗∗∗∗P < 0.0001. TLR4, toll-like receptor 4; PIEZO1, Piezo-type mechanosensitive ion channel component 1; TRPA1, transient receptor potential cation channel subfamily A member 1; LPS, lipopolysaccharide; PBS, phosphate buffer saline; 5-HT, 5-hydroxytryptamine.
Credit
Ruifang Luo, Yuan Miao, Riqiang Hu, Fang Lin, Junyan Yan, Ting Yang, Lu Xiao, Zhujun Sun, Yuting Wang, Jie Chen
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