Fig. 5.

C2IN-C3lim inhibits the 3-dimensional extravasation of primary human monocytes ex vivo. Primary human monocytes were isolated from healthy donors and 3 × 10⁶ cells per sample were incubated at 37 °C for 1 h in RPMI1640 medium without FCS with or without C2IN-C3lim (2 µg/mL), C2I (1 µg/mL) + C2IIa (2 µg/mL) and C2IN-C3lim (1 µg/mL) + C2IIa (2 µg/mL). The cells were applied into the left chamber and exposed towards the chemoattractant fMLP (100 nM) from the right chamber. The chambers were connected by seven 5 × 5 µm wide and 100 µm long channels. Chemotaxis was investigated by microscopic monitoring of cells every 5 s for 1 h and subsequent cell tracking was performed to determine cell velocity (also see supplement movie S1 A and B). The massive dots in each group represent single measured cells. In each group the measurements from five independent preparations were pooled. For each group the total cell number is indicated. Values are given as mean ± SD. Significance was tested using Kruskal–Wallis test with Dunn’s multiple comparisons test (****p < 0.0001)