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. 2014 Feb 18;5(18):8211–8222. doi: 10.18632/oncotarget.1788

Figure 6. MY7-mediated internalization of CD13 and ADAM17; CD13/ADAM17 association.

Figure 6

(A) U937 cells were pulsed with anti-CD13 MY7-PE, anti-ADAM17-PE or mIgG1-PE (negative control) at 4°C, then cultured for 2, 4 and 6 h before analysis for total fluorescence. In separate samples, cells were cultured with MY7 (or its isotype IgG1) for the same times periods, then stained with anti-CD13 MY7-PE, anti-ADAM17-PE or mIgG1-PE (negative control) and analyzed for surface CD13 and ADAM17 expression. Results are expressed as the percentage of the mean fluorescence intensity in MY-7 treated cells vs IgG1-treated cells. (B) U937 cells were lysed, and the lysates were immunoprecipitated with either mIgG1 (lane 2), anti-CD13 (MY7) (lane 3) or anti-ADAM17 (R&D1362) (lane 4). Immunoprecipitates (IP) were washed and separated by SDS/PAGE under reducing conditions and immunoblotted with anti-CD13 (SJ1D1) or anti-ADAM17 (Cell Signalling 3976) to evaluate CD13/ADAM17 interaction. The whole cell lysate served as control.