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. Author manuscript; available in PMC: 2018 Oct 1.

Published in final edited form as: Atherosclerosis. 2017 Sep 9;265:231–241. doi: 10.1016/j.atherosclerosis.2017.09.008

LPS in combination with PA increases adhesion molecule and IL-6 expression in HAECs.

(A–C) HAECs were treated with 0.5 ng/ml of LPS, 100 μM of PA or LPS plus PA for 24 h and ICAM-1 mRNA (A), VCAM-1 mRNA (B) and E-selectin mRNA (C) were quantified using real-time PCR. (D) Immunoblotting of ICAM-1 and VCAM-1 in cytoplasma and membrane. HAECs were treated as described above and the ICAM-1 and VCAM-1 protein in cytoplasma and membrane was detected using immunoblotting. GAPDH and β-actin were also detected as control. € Quantification of membrane ICAM-1 (n=2). (F and G) Time courses of IL-6 mRNA expression (F) and IL-6 secretion (G). HAECs were treated with 0.5 ng/ml of LPS, 100 μM of PA or LPS plus PA for 2, 4, 8, 12, 24, 36 and 48 h. At each time point, total RNA was isolated from cells and culture medium was collected for quantification of IL-6 mRNA using real-time PCR and secreted IL-6 protein using ELISA, respectively. The data (mean ± SD) presented are representative of 3 experiments with similar results.

LPS in combination with PA increases adhesion molecule and IL-6 expression in HAECs.

(A–C) HAECs were treated with 0.5 ng/ml of LPS, 100 μM of PA or LPS plus PA for 24 h and ICAM-1 mRNA (A), VCAM-1 mRNA (B) and E-selectin mRNA (C) were quantified using real-time PCR. (D) Immunoblotting of ICAM-1 and VCAM-1 in cytoplasma and membrane. HAECs were treated as described above and the ICAM-1 and VCAM-1 protein in cytoplasma and membrane was detected using immunoblotting. GAPDH and β-actin were also detected as control. € Quantification of membrane ICAM-1 (n=2). (F and G) Time courses of IL-6 mRNA expression (F) and IL-6 secretion (G). HAECs were treated with 0.5 ng/ml of LPS, 100 μM of PA or LPS plus PA for 2, 4, 8, 12, 24, 36 and 48 h. At each time point, total RNA was isolated from cells and culture medium was collected for quantification of IL-6 mRNA using real-time PCR and secreted IL-6 protein using ELISA, respectively. The data (mean ± SD) presented are representative of 3 experiments with similar results.