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

Figure 2.

Figure 2

Analysis of the folding status of the LDLR mutants: Immunoblot analysis of total cell lysates from cells transiently transfected with HA‐tagged wild‐type or mutant LDLRs, under nonreducing conditions. (A) Immunoblots probed against HA antibody, showing difference in the migration of the mature (upper band) and precursor (lower band) forms of LDLR, among the wild‐type and mutants. (B) Immunoblots probed with LDLR C7 monoclonal antibody that specifically recognizes properly folded, mature LDLR. (C) Endo H susceptibility of the wild‐type LDLR and its mutants: HA‐tagged wild‐type LDLR or mutant variants were transiently expressed in HEK‐293T cells. HA‐tagged proteins were immunoprecipitated, treated with Endo H for 4 h at 37 °C (+) or left untreated for 4 h at 37 °C (−), and analyzed by immunoblotting with anti‐HA antibody. The mature form of the receptor was detectable in the immunoprecipitates from the wild‐type and D445E mutant and was resistant to Endo H digestion. ER forms of the wild‐type as well as the mutants were sensitive to Endo H treatment.