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. 2019 Oct 23;9(11):1994–2005. doi: 10.1002/2211-5463.12740

Figure 4.

Figure 4

Association of LDLR mutants with ER chaperones and ERAD factors: (A) Cell lysates were prepared from transiently transfected cells expressing either wild‐type or mutant LDLR‐HA (D445E, D482H, and C667F). Cell lysates (200 µg) were subjected to Co‐IP using anti‐HA agarose beads. Western blot analysis of the immunoprecipitated complexes was performed using antibodies against GRP78 (BiP), CANX, GRP94, and ERP72. (B) The immunoblots from A were stripped and reprobed with antibodies against OS9, HRD1, SEL1L, and HA. The ER‐retained mutants show stronger interaction with the ERAD factors than the wild‐type and the D445E mutant. The experiments were performed twice for each construct with identical results.