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. 2015 Mar 3;10(3):e0118473. doi: 10.1371/journal.pone.0118473

Fig 2. S1P up-regulates ICAM-1 expression via S1PR1 and S1PR3 in HPAEpiCs.

Fig 2

(A) The mRNA expression of various S1P receptors on HPAEpiCs was determined by RT-PCR. (B) Cells were pretreated with W123 or CAY10444 for 1 h, and then incubated with S1P for 16 h. The ICAM-1 protein expression was determined by Western blot. (C) Cells were pretreated with W123 (10 M) or CAY10444 (10 M) for 1 h, and then incubated with S1P for 4 h. The ICAM-1 mRNA expression and promoter activity were determined by real-time PCR and promoter assay, respectively. (D) Cells were pretreated with W123 (10 M) or CAY10444 (10 M) for 1 h, and then incubated with S1P for 16 h. The THP-1 cells adherence was measured. (E) Cells were transfected with siRNA of scrambled, S1PR1, S1PR2, or S1PR3, and then incubated with S1P (10 μM) for 16 h. The levels of S1PR1, S1PR2, S1PR3, and ICAM-1 proteins were determined by Western blot. Data are expressed as mean (E) or mean±S.E.M. (B, C, and D) of three independent experiments. *P<0.05; # P<0.01, as compared with the cells exposed to S1P alone (B, C, and D) or transfected with siRNA of scrambled+S1P (E).