Fig. 6: mScarlet tag does not interfere with localization or function of CD63.
From: A live cell reporter of exosome secretion and uptake reveals pathfinding behavior of migrating cells
a Representative images of immunofluorescent staining of LAMP1 (cyan or blue) in HT1080-pHluo_M153R-CD63 cells (n = 210 cells from 28 images from three independent experiments) and HT1080-pHluo_M153R-CD63-mScarlet cells (n = 313 cells from 24 images from three independent experiments). pHluo_M153R-CD63 from a single tag and a dual tag is shown in magenta (the upper raw) and green (the lower raw), respectively. CD63-mScarlet is shown in red. Scale bars, 50 µm. b Percentage of LAMP1 colocalized with tagged fluorescent proteins from (a). Data are presented as median with interquartile range. ***P = 0.0009 by two-sided unpaired Student’s t test. c Representative images of immunofluorescent staining of Rab7 in parental HT1080 cells (n = 343 cells from 27 images from four independent experiments) and HT1080-pHluo_M153R-CD63-mScarlet cells (n = 255 cells from 21 images from three independent experiments). Rab7 is shown in cyan (the upper raw) or in blue (the lower raw). Endogenous CD63 from parental HT1080 is shown in magenta. pHluo_M153R-CD63 and CD63-mScarlet are shown in green and red, respectively. Scale bars, 50 µm. d Percentage of Rab7 colocalized with endogenous CD63 and tagged fluorescent proteins from (c). Data are presented with median with interquartile range. Statistics was tested by two-sided unpaired Student’s t test. e Representative images of immunofluorescent staining of CD63 (blue) for extracellular deposits from HT1080-pHluo_M153R-CD63-mScarlet cells (n = 34 images from three independent experiments). pHluo_M153R-CD63 and CD63-mScarlet are shown in green and red, respectively. Scale bar, 50 µm. f Percentage of tagged fluorescent proteins that colocalized with CD63 from (e). Data are presented with median with interquartile range. ***P < 0.001 by two-sided paired Wilcoxon test. g Percentage of pHluo_M153R colocalized with mScarlet from (e). Data are presented with median with interquartile range. Source data are provided as a Source Data file.
