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. Author manuscript; available in PMC: 2020 Dec 26.
Published in final edited form as: Cell Rep. 2020 Dec 8;33(10):108486. doi: 10.1016/j.celrep.2020.108486

Figure 2. Functional Roles of MCU-EMRE Contact Sites.

Figure 2.

(A) Functional effects of site 1 mutations. EMRE and MCU mutants were expressed in EMRE-KO and MCU-KO cells, respectively. Arrows indicate the addition of Ru360.

(B) Effects of PKP-motif mutations on MCU-EMRE interactions. Immobilized, 1D4-tagged WT MCU was used to pull down indicated EMRE mutants.

(C) Experiments testing the functional role of site 1.

(D) Site 3 and its connection to Y289 (orange) via TMH2.

(E) Functional effects of Y279 mutations. The western blot compares expression levels of these Y279 MCU mutants with WT. Actin serves as loading control.

Data are presented as means ± SEMs. Con, hME with no mutations; hME, an MCU-EMRE fusion protein; IP, elution, analyzed using 1D4 or EMRE antibody; SGG, an EMRE mutant with an SGGSGG linker between TMH0 and the PKP motif; WCL, whole-cell lysate input.