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. Author manuscript; available in PMC: 2012 Apr 1.
Published in final edited form as: Biochim Biophys Acta. 2010 Nov 18;1812(4):565–571. doi: 10.1016/j.bbadis.2010.11.003

Figure 6. TNF-α treatment induces HcrtR2 ubiquitination through a pathway mediated by cIAP-1 and cIAP-2.

Figure 6

Panel A: B35 cells overexpressing the HcrtR2 were cultured in the presence and absence of TNF-α cytokine (2 ng/ml) for 6 hours at 37°C. Cells were lysed, and equivalent amount of whole cell detergent lysates were immunoprecipitated (IP) by anti-HcrtR2 IgG, followed by Western blot analysis with the indicated antibodies. Panel B: B35 cells constitutively expressing the HcrtR2 were transfection with siRNA directed toward cIAP-1 or/and cIAP-2 as described in Materials and Methods 2.4, then cultured in the presence and absence of TNF-α cytokine (2 ng/ml) for 24 hours at 37°C. Cells were harvested, and equivalent amount of whole cell detergent lysates were Western blotted with the indicated antibodies. Experiment was repeated three times, and representative images are shown.