Skip to main content
. Author manuscript; available in PMC: 2023 Dec 8.
Published in final edited form as: Nat Cardiovasc Res. 2023 Aug 31;2:835–852. doi: 10.1038/s44161-023-00325-8

Fig. 5. Inhibition of de novo phospholipid biosynthesis pathways leads to reduced MK polyploidization and proplatelet formation.

Fig. 5

a, Schematic representation of FA uptake and phospholipid biosynthesis pathways. Acyl-CoA is generated from FAs via long-chain acyl-CoA synthetase (ACSL) and transferred into glycerol-3-phosphate (G-3-P) by glycerol-3-phosphate acyltransferase (GPAT) to create LPA. Inhibitors of the enzymatic steps are illustrated by red inhibitory arrows. Dotted arrows connect lipids to anionic lipid shift illustrating putative functions in proplatelet formation.

b, Relative amount of depicted lipid classes of control and inhibitor treated MKs (Triacsin C, FSG67) on day 7. Non-treated day 0 MKs were used as baseline control and lipid quantities were set to 1 and the standard deviation is depicted as dotted lines. Changes for all lipid classes were calculated as ratios relative to control day 0.

c, Number of proplatelet-forming cells in vitro of control and inhibitor-treated MKs examined from day 4 for 30 hours (means±SD, n=5).

d, Representative images of proplatelet forming MKs after 24hrs examination.

e, Representative ploidy histogram and arithmetic means of DNA content of control and inhibitor treated CD41+ BM-derived MKs cultured for 5 days (n=6).

f, Representative confocal microscopy images of immunostained femora cryosections (left) and arithmetic means±SD (n=6, right) of MK localization (distance to sinusoids) and fragmentation in the BM of mice subjected to DMSO, Triacsin C, or FSG67 injections for seven days. Green, MKs (GPIb); red, sinusoids (CD105); blue, nuclei (DRAQ5). Asterisks indicate fragmented MKs.

g, Representative ploidy histogram and arithmetic means of DNA content of BM MKs from Triacsin C, FSG67 or vehicle treated mice (n=5-7).

h, Representative median projections of 25 μm z-stacks (left) and arithmetic means±SD (n=5-7, right) of MK fragmentation and proplatelet forming MKs from the BM of Triacsin C, FSG67 or vehicle treated mice. Arrows indicate proplatelets and asterisks indicate fragmented MKs. Green, MKs/platelets (GPIX derivative); magenta, vessels (CD105). Graphs represent ratios of proplatelet-forming (left panel) and fragmented MKs (4-8 z-stacks per mouse).

i, Platelet count of Triacsin C, FSG67 or vehicle treated mice (n=9).

A two-sided t-test was used for statistical analysis (*P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001).