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. 2010 Jul 12;190(1):129–141. doi: 10.1083/jcb.200912087

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© 2010 Kuang et al.

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Figure 3. — iNOS clearance is reduced after stimulus in SPSB2-deficient macrophages. (A and B) BMDMs from either wild-type littermates (Spsb2^+/+) or SPSB2-deficient mice (Spsb2^−/−) were incubated with IFN-γ and LPS for the times indicated (A) or incubated with or without (−) IFN-γ and LPS overnight, washed, replenished with fresh medium, and lysed at the indicated times after wash (B). Lysates were then separated by SDS-PAGE and analyzed by Western blotting using anti-iNOS antibody (top). Equivalent protein loading was confirmed by stripping and reprobing membranes with antitubulin antibody (anti-Tub; bottom). (C) Thioglycollate-elicited peritoneal macrophages from Spsb2^−/− and littermate control mice (Spsb2^+/+) were cultured overnight in medium containing LPS/IFN-γ, washed, replenished with fresh medium, and lysed at the indicated times after wash. iNOS expression was detected by Western blotting (top). Equivalent protein loading was confirmed by stripping and reprobing membranes with antitubulin antibody (bottom).