Figure 3

Disruption of endothelial cell homeostasis in the KD group. (a) HCAECs were treated with medium containing 15% sera from HC (n = 17), CAL- and IVIG-responsive KD (n = 25), CAL+ KD (n = 7), IVIG-resistant KD (n = 11) and convalescence KD (n = 13) subjects. After 24 hours and 48 hours, the proliferation of HCAECs were detected by CCK8 (all samples were normalized to HCAECs with 15% FBS treatment and each sample was examined in triplicate). (b) HCAECs were treated with medium containing 15% sera from HC (n = 3) and KD (n = 8) subjects. Cells proliferation was monitored for 140 h by RT-CES system. Green curve and red curve represent mean cell numbers of KD group and HC group respectively. SR CI indicates slope rate cell index; 85h-CI indicates cell index at 85 h. (c) In vitro angiogenesis. HCAECs were plated on growth factor-reduced Matrigel to migrate or join together in the presence of 15% sera from HC (pooled from 8 individuals) and KD (pooled from 12 individuals) subjects. After 4 hours, cells were photographed at a magnification of 40×. The lower panel of (c) is the statistical analysis of angiogenesis. (d) HCAECs were treated with medium containing 15% sera from HC (n = 8) and KD (n = 29) subjects. After 6 hours, E-selectin, VCAM-1, TF and MCP-1 mRNA levels were measured by qPCR (all samples were normalized to HCAECs with 15% FBS treatment). The bar graphs are mean ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001 and ****P < 0.0001 compared with respective control group.