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. 2021 Apr 28;220(7):e202009179. doi: 10.1083/jcb.202009179

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© 2021 Venkatesan et al.

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Figure S4. — Influences of Mpl and CRT_Del52-D165K and CRT_Del52-D166K mutations upon CRT_Del52 multimerization. (A–C) HEK293T cells were transiently transfected with plasmids encoding CRT_Del52 alone or in combination with His-FLAG Mpl or His-FLAG Mpl alone (A and B) or untagged full-length CRT_Del52, CRT_Del52-D165K, CRT_Del52-D166K, or CRT_Del52-3CA (CRT_{Del52(C163A/C400A/C404A)}) or a control vector (Vec; C). Cell lysates from indicated cells were separated by SDS-PAGE under reducing (8% gels) or nonreducing (4–20% gradient gels) conditions or by native-PAGE (8% gels) and immunoblotted with anti-FLAG or anti-CRT(C_mut) antibodies. In blots following SDS-PAGE under nonreducing and native conditions, species consistent with the size of mutant CRT monomers, dimers, and multimers are indicated. Location of loading wells is also indicated. Data are representative of two sets of analyses. MW, molecular weight.