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. Author manuscript; available in PMC: 2008 Sep 18.
Published in final edited form as: J Biol Chem. 2007 Jan 2;282(9):6183–6191. doi: 10.1074/jbc.M608863200

FIGURE 8. Absence of global ER phenotypes in thyrocytes overexpressing ERp72.

FIGURE 8

A, rtTA-7-ERp72 cells, either uninduced or induced to overexpress ERp72 by 96 h of doxycycline (Dox) treatment, were pulse-labeled with 35S-labeled amino acids for 30 min and chased for the indicated times. At each chase time, the total labeled Tg recovered by immunoprecipitation was either undigested or digested with endoglycosidase H (Endo H). The samples were analyzed by SDS-PAGE and phosphorimaging. B, using a protocol similar to that for described for A, labeled Tg recovered from cells (Lysate) or media were analyzed as a function of chase time. C and D, rtTA-7-ERp72 cells, either uninduced or induced to overexpress ERp72 by 96 h of doxycycline treatment, were treated with tunicamycin continuously for 24 h. At different times of treatment cells were pulse-labeled with 35S-labeled amino acids for 30 min without chase, and newly synthesized Tg was immunoprecipitated and analyzed by SDS-PAGE (C); or cells were lysed and analyzed by Western blotting for ER chaperones ORP150, ERp72, GRP94, or BiP as shown (D).