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. 2020 Oct 22;117(45):27980–27988. doi: 10.1073/pnas.2008885117

Fig. 6.

Fig. 6.

The Mcl-1 TMD translocates the Bok TMD to the mitochondria. (A) Heterooligomerization analysis of the Mcl-1 WT TMD and single amino acid variants with the Bok TMD measured by BiFC in HCT116 cells. (B) Left shows the average structures of Mcl-1–Bok TMD heterodimers based on cluster 2 structures. Right shows the corresponding average amino acid–amino acid occupancy of contact. (C) Confocal images of HeLa cells stained with red ER tracker (Left) or expressing MtDsRed marker (Right) and transfected with VN and VC Bok TMD constructs. Oligomers were observed in the green channel. Merge panels show colocalizations with the PCC. (D) Confocal images of HeLa cells expressing MtDsRed marker and transfected with the VN Mcl-1 TMD plus the VC Bok TMD constructs. As above, oligomers were observed in the green channel. (E) Graph shows average PCC in the mitochondria in C and D experimental conditions. Bars represent mean ± SEM (n = 86 cells per condition); P value, according to Student t test, displayed ***P < 0.001. (F) Representative confocal images and quantification of in situ PLA targeting VDAC1 and IP3R interactions in HeLa cells transfected with VN + VC ΔTMD constructs or VN Mcl-1 TMD and VC Bok TMD as indicated. The experiment was performed three times; error bars represent the mean ± SEM; ***P < 0.001.