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. 2019 Feb;1866(2):264–276. doi: 10.1016/j.bbamcr.2018.11.004

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© 2018 The Authors

This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).

PMC Copyright notice

Fig. 6 — I942 inhibits monocyte adhesion to HUVEC monolayers.

a) Confluent HUVEC monolayers were stimulated for 48 h with IL6 (5 ng/ml) plus sIL6Rα (25 ng/ml) in the presence or absence of blocking antibodies to ICAM1, VCAM1 and SELE. Monolayers were then incubated with fluorescently labelled monocytes and the amount of monocyte attachment to monolayers was calculated using a fluorometer, as described in Materials and methods. The relative degree of monocyte adhesion is with significant increases in adhesion being indicated, *, p < 0.05, and significant decreases in adhesion relative to IL6 treated cells with control antibody are also indicated ^#, p < 0.05 (n = 3). Images of adherent monocytes are shown in the lower panel.

b) HUVECs were stimulated with IL6 (5 ng/ml) plus sIL6Rα (25 ng/ml) in the presence or absence of 100 μM I942 or 50 μM 007. Increases in monocyte adhesion are indicated, *, p < 0.05, as are significant decreases relative to IL6-stimulated cells, ^#, p < 0.05 and ^###, p < 0.001 (n = 3). Non-significant changes are also indicated (ns). Images of adherent monocytes are shown in the lower panel.

c) In the upper panel, confluent HUVEC monolayers were stimulated for 48 h with TNFα (10 ng/ml) in the presence or absence of blocking antibodies to ICAM1, VCAM1 and SELE. Monolayers were then incubated with fluorescently labelled monocytes and the amount of monocyte attachment to monolayers was calculated using a fluorometer, as described in Materials and methods. The relative degree of monocyte adhesion is with significant increases in adhesion being indicated, **, p < 0.01 and ***, p < 0.001, and significant decreases in adhesion relative to TNFα treated cells with control antibody are also indicated ^###, p < 0.001 (n = 3). In the lower panel, HUVECs were TNFα (10 ng/ml) for 5 h in the presence or absence of 100 μM I942. Increases in monocyte adhesion are indicated, ^###, p < 0.001 (n = 3), as are significant decreases relative to TNFα -stimulated cells, ^###, p < 0.001.

Fig. 6 — I942 inhibits monocyte adhesion to HUVEC monolayers.

a) Confluent HUVEC monolayers were stimulated for 48 h with IL6 (5 ng/ml) plus sIL6Rα (25 ng/ml) in the presence or absence of blocking antibodies to ICAM1, VCAM1 and SELE. Monolayers were then incubated with fluorescently labelled monocytes and the amount of monocyte attachment to monolayers was calculated using a fluorometer, as described in Materials and methods. The relative degree of monocyte adhesion is with significant increases in adhesion being indicated, *, p < 0.05, and significant decreases in adhesion relative to IL6 treated cells with control antibody are also indicated ^#, p < 0.05 (n = 3). Images of adherent monocytes are shown in the lower panel.

b) HUVECs were stimulated with IL6 (5 ng/ml) plus sIL6Rα (25 ng/ml) in the presence or absence of 100 μM I942 or 50 μM 007. Increases in monocyte adhesion are indicated, *, p < 0.05, as are significant decreases relative to IL6-stimulated cells, ^#, p < 0.05 and ^###, p < 0.001 (n = 3). Non-significant changes are also indicated (ns). Images of adherent monocytes are shown in the lower panel.

c) In the upper panel, confluent HUVEC monolayers were stimulated for 48 h with TNFα (10 ng/ml) in the presence or absence of blocking antibodies to ICAM1, VCAM1 and SELE. Monolayers were then incubated with fluorescently labelled monocytes and the amount of monocyte attachment to monolayers was calculated using a fluorometer, as described in Materials and methods. The relative degree of monocyte adhesion is with significant increases in adhesion being indicated, **, p < 0.01 and ***, p < 0.001, and significant decreases in adhesion relative to TNFα treated cells with control antibody are also indicated ^###, p < 0.001 (n = 3). In the lower panel, HUVECs were TNFα (10 ng/ml) for 5 h in the presence or absence of 100 μM I942. Increases in monocyte adhesion are indicated, ^###, p < 0.001 (n = 3), as are significant decreases relative to TNFα -stimulated cells, ^###, p < 0.001.