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. 2015 Dec 9;5:18007. doi: 10.1038/srep18007

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Copyright © 2015, Macmillan Publishers Limited

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(a) Dose dependent regulation of A20, IRAK1 and TRAF6 protein levels by IL-1β. HMC cells were treated with the indicated concentrations of IL-1β for 2 h. The protein levels of A20 (82 KD), IRAK1 (78 KD), and TRAF6 (60 KD) were detected by western blot. GAPDH (36 KD) protein levels were detected as loading controls. (b) Time-dependent regulation of A20, IRAK1, and TRAF6 protein levels by IL-1β. (c) Dose-dependent regulation of IRAK1 and TRAF6 mRNA levels by IL-1β. (d) Time dependent regulation of A20, IRAK1 and TRAF6 protein levels by IL-1α. (e) The effect of A20 over-expression on the protein levels of A20, IRAK1 and TRAF6. HMC cells, transfected with 2 μg plasmid containing the sequence coding for human A20 protein, or with empty plasmid, G418-resistant clones were selected for the detection of A20, IRAK1, and TRAF6 expression by western blot. GAPDH protein levels were detected as loading controls. (f) The effect of A20 down-regulation on the protein levels of A20, IRAK1, and TRAF6. HMC cells, transfected with the indicated concentrations of A20 siRNA, or control siRNA, for 5 h, were cultured for 36 h, followed by the treatment with 5 ng/ml IL-1β for 2 h. The protein levels of A20, IRAK1, and TRAF6 were detected by western blot. GAPDH protein levels were detected as loading controls.