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. 2020 Oct 30;11:569802. doi: 10.3389/fphar.2020.569802

FIGURE 3.

FIGURE 3

S1P induces COX-2 expression via a Gq- or PTX-sensitive Gi protein-coupled S1PR1/3. (A,B) Cells were pretreated with GPA2A (Gq antagonist) for 1 h or pertussis toxin (PTX, Gi protein inactivator) for 24 h and then incubated with 15 μM S1P for 8 h. (C) Cells were transfected without or with COX-2 promoter-luciferase reporter gene, pretreated with GPA2A (1 μM) or PTX (100 ng/ml) for 1 h, and then incubated with 15 μM S1P for 4 h (mRNA) or 1 h (promoter). The COX-2 mRNA and promoter activity were analyzed by real time-PCR (open bar) and promoter assay (gray bar). (D) Cells were transfected with siRNA for scrambled, Gqα, or Giα for 24 h and then exposed to 15 μM S1P for 8 h (A,B,D) The levels of COX-2, GAPDH, Gqα, and Giα proteins were analyzed by Western blot. Data are expressed as mean ± SEM of three individual experiments (n = 3). # p < 0.05, as compared with the cells treated with S1P alone.