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. Author manuscript; available in PMC: 2018 Jul 1.
Published in final edited form as: J Immunol. 2017 May 24;199(1):263–270. doi: 10.4049/jimmunol.1600409

Figure 5.

Figure 5

Confocal microscopy reveals that SLC46A2-EGFP clusters in response to TCT. HEK-293T cells were transfected with SLC46A2-EGFP, and exposed to LysoTracker (magenta). (A) mSlc46a2-EGFP (green) is evenly distributed and co-localized with LysoTracker-positive organelles throughout the cytoplasm without TCT. 2 representative images from independent assays, exhibiting the range of patterns of observed. (B) After 30-minute exposure to TCT, mSlc46a2-EGFP-expressing, LysoTracker-positive organelles relocalized to form large aggregates. (C) Sizes of EGFP+ and Lysotracker+ puncta were measured and compared in the absence of TCT (n=110 puncta from of 13 cells) or presence of TCT (n=129 puncta from 21 cells) from two independent assays. (D) Human SLC46A2-EGFP protein showed similar behavior as its mouse homolog upon TCT exposure (n=137 puncta from 30 cells for vehicle, n=173 puncta from 35 cells for TCT). All data points are presented, and the mean is indicated. ** p < 0.01, *** p < 0.001, unpaired two-tailed t-test. For all images, the scale bar is 5 μm.