Fig. 5. Lack mtCU interaction does not affect the MICU1-MIC60 interaction.

(A) Schematic representation of MICU1 domains and the point mutants utilized in the present study. (B) Micu1^−/− MEFs were reconstituted with Flag-tagged MICU1 WT, MICU1 EMRE^mut, MICU1 MCU^mut, MICU1 mtCU^mut, MICU1 EF1^mut, MICU1 EF2^mut, MICU1 EF1/2^mut, and MICU1 dimer^mut mutants. FLAG immunoprecipitates were probed for FLAG, MIC60, and MCU to detect MICU1-MIC60 and MICU1-MCU interactions. Western blots are representative of 3 independent experiments. (C) Schematic for the MICU1 truncation mutants utilized in the present study. (D) Flag-tagged MICU1 WT, T1, T2, T3, T4, and T5 mutants were reconstituted in Micu1^−/− MEFs. FLAG immunoprecipitates were probed for FLAG and MIC60 to detect MICU1-MIC60 interactions. Western blots are representative of 3 independent experiments.