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. 2014 Jun 2;9(6):e98606. doi: 10.1371/journal.pone.0098606

Figure 4. WASH-deficiency in DCs leads to polyubiquitination of MHCII and its accelerated degradation.

Figure 4

(A) BMDCs from WASHf/f Vav-Cre and control Vav-Cre mice were antibody stained for surface MHCII and analyzed using flow cytometry. Histograms and the bar graph are representative of six separate experiments. (B) WASHf/f LysM-Cre mice and control LysM-Cre mice BMDCs were surface labeled with biotin and chased in culture for the indicated time points. After chase, cells were lysed to immunoprecipitate MHCII. Immunoprecipitates were then resolved by PAGE and processed by immunoblot to detect remaining biotinylated MHCII with streptavidin (SAv). (C) Decay of surface labeled MHCII over time was measured by densitometry of immunoblots. (D) Percent MHCII recycling was measured on WASHf/f Vav-Cre and control Vav-Cre BMDCs using acid stripping as described in the Materials and Methods. Data are representative of three separate experiments. Bars represent mean ≥ SEM, *p≤0.05.