Fig.6. IVIG and plate-bound IgG regulate RANKL-induced signaling pathways, in part, by inducing A20 expression.

(a-b) Human monocytes were cultured for one day with human M-CSF (20 ng/ml) in control wells, wells with 1 mg/ml of IVIG, or wells coated with 100 µg/ml of IgG (labeled X-linked). Cells were harvested at 0, 3, 6 and 24 hours after stimulation. (a) mRNA was measured using RT-qPCR and normalized relative to the expression of GAPDH. (b) Immunoblot of whole cell lysates with A20 and p38 antibodies. Representative images from three independent donors. (c) 40 ng/ml RANKL was added for the indicated times, and cells were harvested. Immunoblot of whole cell lysates using I-κBα and p38 antibodies. Representative data from at least three independent experiments are shown. (d) OCPs were treated with 40 ng/ml RANKL for one hour and nuclear lysates were immunoblotted with p50/p105, p52/p100, p65, and lamin B antibodies. Representative data from four independent donors.