Fig. 3: VCAM-1 regulates IL-33-induced IL-8 expression in HRMVECs.

a HRMVECs were transfected with control siRNA (siControl) or VCAM-1 siRNA (siVCAM-1), quiesced, treated with or without IL-33 (20 ng/mL) for 8 h, cell extracts were prepared and equal amount of protein from the mentioned groups were analysed for angiogenic genes using proteome profiler human angiogenesis array kit. b, c Quiescent HRMVECs were treated with IL-33 (20 ng/mL) for the indicated time periods and total cellular RNA and cell extracts were prepared and analysed by QRT-PCR and western blotting, respectively for IL-8 expression. d, e HRMVECs were transfected with control siRNA (siControl) or VCAM-1 siRNA (siVCAM-1), quiesced, total cellular RNA and cell extracts were prepared and analysed by QRT-PCR for IL-8 and by western blotting for the indicated proteins. f, g Quiescent HRMVECs were first treated with QNZ (10 μM, NFκB inhibitor) or PDTC (10 μM, NFκB inhibitor) for 30 min, followed with or without IL-33 (20 ng/mL) treatment for 8 h. The cell lysates were prepared and analysed for IL-8 levels by Western blotting and normalized to β-tubulin. n = 3 (b, d) or n = 4 (c) biologically independent samples per group, expressed as Mean ± SD. *P < 0.05 vs control or siControl, **p < 0.05 vs siControl + IL-33.