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. 2011 Mar 24;286(19):16929–16939. doi: 10.1074/jbc.M110.215871

FIGURE 5.

FIGURE 5.

Effect of transiently expressed HRD1-SEL1L on the degradation of NHK and NHK-QQQ. A, cells transfected with NHK, SEL1L, and/or HRD1 were pulse-labeled for 15 min and chased for the times indicated. Aliquots of cells extracted by 3% digitonin were immunoprecipitated (IP) with anti-α1 antitrypsin (α1AT) antibody. The arrowhead indicates the position of NHK. The open arrow shows coimmunoprecipitated HRD1myc. B, shown is quantification of NHK degradation in A. The relative radioactivity of NHK was calculated by setting the NHK intensity at the end of the pulse time as 100%. Results are the mean and S.E. (n = 3). C, cells transiently expressing NHK-QQQ, SEL1L, and/or HRD1 were metabolically labeled, and the degradation kinetics of NHK-QQQ was analyzed as in A. The arrowhead indicates the position of NHK-QQQ. D, radioactivity of NHK-QQQ in C was quantified as in B. E, cells were transfected with NHK-QQQ and SEL1L and treated with or without MG132 for 4 h before metabolic labeling. Arrowheads and brackets indicate the positions of NHK-QQQ and SEL1L, respectively. Asterisks are the degradation products generated from transiently expressed SEL1L.