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. Author manuscript; available in PMC: 2019 Mar 6.
Published in final edited form as: Biochem J. 2018 Mar 6;475(5):873–886. doi: 10.1042/BCJ20170884

Figure 5. Expression of FVIII missense mutants induces BiP and increases interactions with BiP and CRT.

Figure 5

(A) Thirty-six hours after transfection of the indicated mutants, expression levels in input cell extracts were analyzed by immunoblotting with the indicated antibodies. Interactions between A2 mutants and ER chaperones were analyzed by co-immunoprecipitation (IP). Cell extracts were immunoprecipitated with a mouse monoclonal anti-FLAG antibody or normal mouse IgG. The immunoprecipitates from anti-FLAG IP and IgG control IP were analyzed by immunoblotting with the indicated antibodies. CRT: calreticulin; CNX: calnexin. Arrow points to the CRT band in the IP data. The lower band is non-specific as it also appears in IgG controls. (B) The amounts of BiP in cell lysates, BiP and CRT that co-immunoprecipitated with A2 domain mutants were densitometrically quantified and expressed as fold changes relative to WT. Data are mean ± SEM, n = 3, *P < 0.05.