Translational events in platelets are increased coordinately in clinical and experimental sepsis. (A) Isolated platelets from septic patients or matched, healthy donors were incubated with ³⁵S-methionine (t = 18 hours) to label newly synthesized proteins and then analyzed by 2D gel electrophoresis. Representative ³⁵S-methionine–labeled 2D gel of platelets from a septic patient and healthy donor (representative of n = 3/group). (B) Schematic of protocol for sequencing total RNA (RNA-seq) and ribosome protected mRNA reads (RPRs, ribosome footprint profiling) in isolated human and murine platelets. (C) Ribosome footprint profiling of platelets from healthy donors and septic patients (n = 3/group) demonstrating global triplet periodicity. Increased reads align with frame 0 (red) compared with frame 1 (green) and frame 2 (blue). Frame number indicates distance from the first nucleotide of the P site. (D) Depiction of global number of reads per transcript for total RNA (healthy, green; sepsis, purple) and RPR (healthy, red; sepsis, blue) relative to the start and stop codon. (E) Correlation plot of the expression of orthologous, conserved RPRs in platelets from untreated control mice (n = 4) and healthy human donors (n = 3). (F) Correlation plot of the expression of orthologous, conserved RPRs in platelets from CLP-treated mice (n = 3, pooled platelets) and septic patients (n = 3).