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. Author manuscript; available in PMC: 2011 Dec 22.
Published in final edited form as: Mol Cell. 2010 Dec 22;40(6):917–926. doi: 10.1016/j.molcel.2010.11.033

Figure 2. NS-1 κ LC is not ubiquitylated at the N-terminus.

Figure 2

(A) COS-1 cells expressing either WT NS-1 LC or the Xa4 mutant were treated with MG132 for 5 hr and isolated LC were cleaved with Factor Xa followed by a second immunoprecipitation with an anti-κ LC antibody that recognizes the CL domain. Ubiquitylation of the LC were analyzed by immunoblotting. The uncleaved form is indicated with an asterisk and the cleaved form with two asterisks. (B) The indicated untagged LC constructs were expressed in COS-1 cells and lysates were prepared from cells treated with or without lactacystin. LC were isolated and subjected to immunoblot analyses with the mouse anti-Ub antibody. (C) 293T cells expressing the indicated mutants were pulse labeled and chased for the indicated times. Isolated LC were separated by reducing SDS-PAGE, and detected by autoradiography. Percent LC remaining at each time point is indicated under each lane.