Inhibition of RhoA activity in cold-stored platelets improve their function and half-life. (A) Upon cold storage of platelet concentrates, platelets are activated, and stress fibers are formed, mediated by RhoA activity, leading to consequent downstream signaling, apoptosis, and clearance. β-GlcNAc glycan residues become exposed on the platelet surface, acting as an “eat-me” signal, recognized by Ashwell-Morell receptors on hepatocytes in the liver. The addition of Rho inhibitors prevents stress fiber formation and all major activation processes, as per panel B, preventing rapid platelet clearance. (B) After cold-storage of platelet concentrates, GPIbα reorganization within the membrane leads to 14-3-3ζ recruitment, release of procoagulant phosphatidylserine (PS)⁺ extracellular vesicles (EVs), mitochondrial depolarization, and death. In addition, several glycan-modulating enzymes are translocated to signaling platforms, specifically the lipid rafts, including UDP N-acetyl-D-galactosamine (GalNAc):polypeptide N-acetyl-galactosaminyltransferases T1 and T3 (GalNAc-T1 and -T3), sialyltransferase, and UDP-galactosyltransferase. GDP, guanosine diphosphate. Claire Linnane (Australian Red Cross Lifeblood) provided some of the building blocks of this cartoon.