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. 2019 Jan 7;218(1):251–266. doi: 10.1083/jcb.201802157

Figure 1.

Figure 1.

In U2OS cells, JMY translocates from STRAP-positive nonmotile foci to motile LC3-positive motile foci upon starvation. (A) JMY (red) and STRAP (blue) colocalize on foci under normally fed conditions (top). More JMY (red) colocalizes with LC3 (green) when cells are starved in HBSS (bottom). (B) Two vesicles (arrowheads) that contain both JMY and LC3 but no STRAP (yellow color in merge channel) move persistently through cytoplasm. Kymographs show travel distance versus time. (C) Enlarged box indicates two populations of vesicles. (1) Vesicles that contain JMY and STRAP but no LC3 (magenta color in merge channel) are nonmotile. Kymograph 1 shows one example of a nonmotile JMY- and STRAP-positive vesicle. (2) Vesicles that contain all three proteins, JMY, STRAP, and LC3 (yellow-white color in merge channel), move in a saltatory manner. Kymograph 2 shows one example of the saltatory movement of a JMY-, STRAP-, and LC3-positive vesicle. (D) JMY- and LC3-positive vesicle (yellow color in merge channel, arrowhead) moves and fuses with JMY- and STRAP-positive (magenta color in merge channel) vesicle. The fused vesicle is nonmotile during observation. (E) Quantification of colocalization of JMY, STRAP, and LC3 under fed and starved conditions. The percentage is normalized to total number of puncta (including all LC3, JMY, and STRAP puncta) of the corresponding cell (n = 3 independent experiments, 24 cells, total number of puncta per cell: 15 ~ 83). Quantifications of colocalization normalized to each protein are shown in Fig. S1 (A–C). (F) Quantification of migration speed of JMY-positive vesicles (number of JMY- and LC3-positive puncta: 73; number of JMY- and STRAP-positive puncta: 208; number of JMY-, LC3-, and STRAP-positive puncta: 73, n = 3, **, P < 0.01, Student’s t test. For all box plots: boxes represent upper and lower quartile with median value displayed as a line; the lines extending from each box mark the minimum and maximum of data set within acceptable range; the range is defined as quartile limit ±1.5 (*, interquartile distance). Temperature: 37°C. Scale bars: whole cell, 5 µm; zoom-in box, 2 µm.