Figure 4
Figure 4. Loss of RanBP10 results in a disordered platelet marginal band. Platelets were obtained from adult mice by retro-orbital blood withdrawal, fixed, and subsequently analyzed by transmission electron microscopy. (A) The overview of wild-type platelets showed the typical discoid shape of platelets with normal granular content. (B) RanBP10−/− platelets exhibit no differences in granular content but a less discoid shape. (C) The change of the platelet shape was quantified by determination of the geometric axis ratio, which shows a statistical significant increase (P < .05; t test) between wild-type (n = 21) and RanBP10-null mice (n = 25). (D) Wild-type platelets possess a normal amount8–12 of MT coils (MT, black arrows) in the periphery. (E) In contrast some RanBP10-deficient platelets exhibit a greater amount (here up to 25) of filaments per coil coils (MT, black arrows). (F) Other platelets showed a normal number of MT filaments per coil, but an atypical appearance in profile (black arrow) and axis (black arrowhead) in one cross-section. (G) RanBP10-knockout platelets also showed additional MT coils (MT, black arrow). Immunofluorescence analysis was consistent with electron microscopy regarding the marginal band. (D) Inset shows wild-type platelets immunostained with an antibody against β-tubulin, with typical MT coils, representing the marginal band. (E) Inset shows the disordered marginal band of RanBP10−/− platelets by immunofluorescence microscopy. Despite having a normal marginal band (not shown), the platelets exhibit multiple starting points of MT coils (white asterisk) and structures that look like halved MT coils (white arrows). (H) Quantification of filaments per MT coil in wild-type (n = 17) and RanBP10 knockout platelets (n = 15) shows a small increase of MT coils for the RanBP10-deficient platelet, but the high standard deviation indicates that the number of MT coils ranges from well above to well below normal. Original magnification, ×10 000 (A-B); ×∼ 35 000 (D-G). Scale bars (insets D-E) indicate 2 μm. The resolved images and quantifications are representatives of 2 independent experiments.

Loss of RanBP10 results in a disordered platelet marginal band. Platelets were obtained from adult mice by retro-orbital blood withdrawal, fixed, and subsequently analyzed by transmission electron microscopy. (A) The overview of wild-type platelets showed the typical discoid shape of platelets with normal granular content. (B) RanBP10−/− platelets exhibit no differences in granular content but a less discoid shape. (C) The change of the platelet shape was quantified by determination of the geometric axis ratio, which shows a statistical significant increase (P < .05; t test) between wild-type (n = 21) and RanBP10-null mice (n = 25). (D) Wild-type platelets possess a normal amount8-12  of MT coils (MT, black arrows) in the periphery. (E) In contrast some RanBP10-deficient platelets exhibit a greater amount (here up to 25) of filaments per coil coils (MT, black arrows). (F) Other platelets showed a normal number of MT filaments per coil, but an atypical appearance in profile (black arrow) and axis (black arrowhead) in one cross-section. (G) RanBP10-knockout platelets also showed additional MT coils (MT, black arrow). Immunofluorescence analysis was consistent with electron microscopy regarding the marginal band. (D) Inset shows wild-type platelets immunostained with an antibody against β-tubulin, with typical MT coils, representing the marginal band. (E) Inset shows the disordered marginal band of RanBP10−/− platelets by immunofluorescence microscopy. Despite having a normal marginal band (not shown), the platelets exhibit multiple starting points of MT coils (white asterisk) and structures that look like halved MT coils (white arrows). (H) Quantification of filaments per MT coil in wild-type (n = 17) and RanBP10 knockout platelets (n = 15) shows a small increase of MT coils for the RanBP10-deficient platelet, but the high standard deviation indicates that the number of MT coils ranges from well above to well below normal. Original magnification, ×10 000 (A-B); ×∼ 35 000 (D-G). Scale bars (insets D-E) indicate 2 μm. The resolved images and quantifications are representatives of 2 independent experiments.

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