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. 2019 Mar 29;10:285. doi: 10.3389/fphys.2019.00285

FIGURE 5.

FIGURE 5

Roles of MAPKs/NF-κB signaling pathway in the cold temperature-induced production of inflammatory cytokines in 16HBE cells. (A) Protein expression of ERK, P38, JNK, and IκBα was analyzed by western blotting in 16HBE cells exposed to medium, 18°C, and 18°C with TRPM8 shRNA transfection or scramble shRNA transefection, respectively. (B) NF-KB activity was measured with luciferase assay in 16HBE cells exposed to medium, 18°C, and 18°C with TRPM8 shRNA transfection or scramble shRNA transefection, respectively. (C) mRNA and protein expression of IL-6, IL-8, and TNF-α were separately measured with RT-RCR and ELISA in 16HBE cells exposed to 18°C with pretreatment with an ERK inhibitor (U0126), a P38 inhibitor (SB203580), or a NF-κB inhibitor (BAY 11-7085; BAY), p- and t- represent phospho- and total-, respectively. Data in each group are mean ± SD n = 3. ∗p < 0.05 vs. control.