Figure 1.
The number of mitochondria per platelet differs between resting and spread platelets. (A) Detection of mitochondria using TOMM20 immunostainings (red). Microtubules were stained using a monoclonal antibody against α-tubulin (cyan). Epifluorescence images of resting (top) and spread platelets (bottom) are shown; scale bar, 10 μm. (B) Detection of mitochondria in platelets incubated for 20 minutes with a mitoTracker (red) before allowing them to spread for 60 minutes on a glass surface and then fixing them (bottom). Resting platelets were fixed in suspension and centrifuged onto PDL-coated coverslips (top). Platelets were then stained for α-tubulin (cyan) and processed for expansion. Maximum intensity projections of confocal image stacks are shown; scale bar, 10 μm, corresponding to 2.5 μm after correction for expansion. (C) Detection of mitochondria in platelets incubated for 20 min with a mitoTracker (red) before allowing them to spread for 60 min on a glass surface and then fixing them (bottom). Resting platelets were fixed in suspension and centrifuged onto PDL-coated coverslips (top). Platelets were then stained for the integrin subunit αIIb (cyan) and processed for expansion. Maximum intensity projections of confocal image stacks are shown; scale bar, 10 μm, corresponding to 2.5 μm after correction for expansion. (D) Quantification of the number of mitochondria in resting platelets after an incubation at 4°C to depolymerize microtubules and in platelets spread for 60 minutes at 37°C on a glass surface using maximal projections of confocal image stacks of unexpanded samples; results for 2 representative donors (D1 and D2) are shown. Data comparison was performed using the 2-tailed Mann-Whitney test. (E) Graph illustrating the number of mitochondria per resting platelet with respect to platelet size (2 representative donors used in panel D). (F) Graph illustrating the number of mitochondria per spread platelet with respect to platelet size (2 representative donors used in panel D). (G) Time course of platelet spreading on a glass surface and quantification of the number of mitochondria per platelet using maximal projections of confocal image stacks of expanded samples. Data comparison was performed using 1-way analysis of variance.

The number of mitochondria per platelet differs between resting and spread platelets. (A) Detection of mitochondria using TOMM20 immunostainings (red). Microtubules were stained using a monoclonal antibody against α-tubulin (cyan). Epifluorescence images of resting (top) and spread platelets (bottom) are shown; scale bar, 10 μm. (B) Detection of mitochondria in platelets incubated for 20 minutes with a mitoTracker (red) before allowing them to spread for 60 minutes on a glass surface and then fixing them (bottom). Resting platelets were fixed in suspension and centrifuged onto PDL-coated coverslips (top). Platelets were then stained for α-tubulin (cyan) and processed for expansion. Maximum intensity projections of confocal image stacks are shown; scale bar, 10 μm, corresponding to 2.5 μm after correction for expansion. (C) Detection of mitochondria in platelets incubated for 20 min with a mitoTracker (red) before allowing them to spread for 60 min on a glass surface and then fixing them (bottom). Resting platelets were fixed in suspension and centrifuged onto PDL-coated coverslips (top). Platelets were then stained for the integrin subunit αIIb (cyan) and processed for expansion. Maximum intensity projections of confocal image stacks are shown; scale bar, 10 μm, corresponding to 2.5 μm after correction for expansion. (D) Quantification of the number of mitochondria in resting platelets after an incubation at 4°C to depolymerize microtubules and in platelets spread for 60 minutes at 37°C on a glass surface using maximal projections of confocal image stacks of unexpanded samples; results for 2 representative donors (D1 and D2) are shown. Data comparison was performed using the 2-tailed Mann-Whitney test. (E) Graph illustrating the number of mitochondria per resting platelet with respect to platelet size (2 representative donors used in panel D). (F) Graph illustrating the number of mitochondria per spread platelet with respect to platelet size (2 representative donors used in panel D). (G) Time course of platelet spreading on a glass surface and quantification of the number of mitochondria per platelet using maximal projections of confocal image stacks of expanded samples. Data comparison was performed using 1-way analysis of variance.

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