Figure 2.
Figure 2. Organelles move bidirectionally in proplatelets. (A-B) Mitochondria move bidirectionally in proplatelets. Mitochondria in megakaryocytes were labeled with MitoTracker, and organelle movements in proplatelets were recorded with fluorescence time-lapse microscopy (Movie S1). Micrographs (A, fluorescence; inset, DIC) show the distribution of mitochondria in a field of proplatelets after labeling with MitoTracker. (B) Kymograph of the boxed region in panel A. Fluorescent images of the labeled mitochondria were taken every 5 minutes. Two mitochondria, highlighted in green, move up in tandem and are observed to separate at 25 minutes. Three mitochondria, highlighted in red, move down during the recording period. The mitochondria pass each other at the 10-minute time point. (C-D) α-granules in proplatelets translocate bidirectionally (Movie S2). The α-granules were labeled by incubating megakaryocytes with Oregon Green 488 fibrinogen conjugate, which is taken up and stored in α-granules. The distribution and dynamics of the labeled α-granules were followed by time-lapse fluorescence microscopy. (C) Micrograph of a proplatelet field labeled with Oregon Green 488 fibrinogen conjugate. (Inset) Imaged differential-interference-contrast micrograph. (D) Kymograph showing time-lapse from the boxed region in panel C. Fluorescent images of the labeled α-granules were taken every 5 minutes. Two α-granules, highlighted in green, come together and move up until one separates (60 minutes) and then moves down (60-75 minutes). An α-granule highlighted in blue remains stationary during the recording period. (E-F) Dense granules move bidirectionally in proplatelets (Movie S3). Megakaryocytes were incubated with mepacrine and washed, and the distribution and dynamics of the dense granules were followed by fluorescence time-lapse microscopy. (E) Fluorescence micrograph of proplatelet field labeled with mepacrine. (Inset) A differential-interference-contrast image of the proplatelet in panel E. (F) Kymograph of the boxed region in panel E. Images of the fluorescently labeled dense granules are every 5 minutes. A group of dense granules, highlighted in blue, remain stationary throughout the recording period. A dense granule, highlighted in green, moves to the left (toward the proplatelet tip) and enters the stationary group. A dense granule, highlighted in red, exits the stationary group and moves to the right toward the base of the proplatelet (55-70 minutes).

Organelles move bidirectionally in proplatelets. (A-B) Mitochondria move bidirectionally in proplatelets. Mitochondria in megakaryocytes were labeled with MitoTracker, and organelle movements in proplatelets were recorded with fluorescence time-lapse microscopy (Movie S1). Micrographs (A, fluorescence; inset, DIC) show the distribution of mitochondria in a field of proplatelets after labeling with MitoTracker. (B) Kymograph of the boxed region in panel A. Fluorescent images of the labeled mitochondria were taken every 5 minutes. Two mitochondria, highlighted in green, move up in tandem and are observed to separate at 25 minutes. Three mitochondria, highlighted in red, move down during the recording period. The mitochondria pass each other at the 10-minute time point. (C-D) α-granules in proplatelets translocate bidirectionally (Movie S2). The α-granules were labeled by incubating megakaryocytes with Oregon Green 488 fibrinogen conjugate, which is taken up and stored in α-granules. The distribution and dynamics of the labeled α-granules were followed by time-lapse fluorescence microscopy. (C) Micrograph of a proplatelet field labeled with Oregon Green 488 fibrinogen conjugate. (Inset) Imaged differential-interference-contrast micrograph. (D) Kymograph showing time-lapse from the boxed region in panel C. Fluorescent images of the labeled α-granules were taken every 5 minutes. Two α-granules, highlighted in green, come together and move up until one separates (60 minutes) and then moves down (60-75 minutes). An α-granule highlighted in blue remains stationary during the recording period. (E-F) Dense granules move bidirectionally in proplatelets (Movie S3). Megakaryocytes were incubated with mepacrine and washed, and the distribution and dynamics of the dense granules were followed by fluorescence time-lapse microscopy. (E) Fluorescence micrograph of proplatelet field labeled with mepacrine. (Inset) A differential-interference-contrast image of the proplatelet in panel E. (F) Kymograph of the boxed region in panel E. Images of the fluorescently labeled dense granules are every 5 minutes. A group of dense granules, highlighted in blue, remain stationary throughout the recording period. A dense granule, highlighted in green, moves to the left (toward the proplatelet tip) and enters the stationary group. A dense granule, highlighted in red, exits the stationary group and moves to the right toward the base of the proplatelet (55-70 minutes).

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