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. 2020 May 5;11:595. doi: 10.3389/fphar.2020.00595

Figure 6.

Figure 6

Conditioned medium of HaCaT cells stimulated with FFA1 agonists increased chemotaxis of neutrophils. (A) Neutrophils were isolated from the blood of healthy donors and analyzed by flow cytometry prior to use in a transwell assay using the conditioned medium of HaCaT cells as a chemoattractant. (B, C) Neutrophil chemotaxis was analyzed using conditioned media from HaCaT cells treated with different concentrations of LA (10–100 μM) for 24 h. The data are represented as the number of neutrophils that migrated to the conditioned medium. The data are the mean ± SEM of five blood donors. *P < 0.05 and **P < 0.01, compared with the control, using Dunnett’s multiple comparison test. (D) Conditioned media of HaCaT treated with 10 µM GW1100 for 15 min and then stimulated with 50 µM LA for 24 h was used to evaluate neutrophil chemotaxis. The data are the mean ± SEM of five blood donors. *P < 0.05 and ***P < 0.001, compared with the control or linoleic acid-treated cells using Tukey’s multiple comparison test. (E, F) Neutrophil chemotaxis was analyzed using conditioned media from HaCaT cells treated with different concentrations of GW9508 (10–100 μM) for 24 h. The data are represented as the number of neutrophils that migrated to the conditioned medium. The data are the mean ± SEM of five blood donors. **P < 0.01, and ****P < 0.0001, compared with the control, using Dunnett’s multiple comparison test.