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. 2019 Mar 15;294(18):7177–7193. doi: 10.1074/jbc.RA118.005659

Figure 1.

Figure 1.

Stably expressed affinity-tagged MSec induces TNT formation in U2OS cells. A, MSec was cloned into a mammalian expression vector that imparts a multiaffinity tag (His-SBP-FLAG) embedded inside a fluorescent (mVenus) tag. B, expression levels of stably expressing empty vector (eMTAP) and exogenous mouse MSec (MSec-MTAP) equivalent to the endogenous human MSec assessed by immunoblotting (IB). C, live confocal microscope images taken from stably expressing MSec-MTAP and empty MTAP cells (both green). TNTs are indicated by arrows. The analyzed TNTs were not adhered to the substratum. Shown is an x-z/y-z visualization of confocal z-stacks of TNTs that were counted in both cell lines (MSec-MTAP and eMTAP). Scale bar, 20 μm. D, quantification of the number of TNTs per 100 cells from the confocal images. Data represent mean ± S.D. (error bars) based on three independent experiments, 100 cells counted per experiment (paired t test, two-tailed; **, p < 0.01). E, time lapse confocal images showing the transport of mitochondria from one cell to another. Arrowheads, position of the mitochondria.