Changes in protein synthesis caused by LDHA deficiency are related to eEF2. (A) Coomassie brilliant blue–stained gel of human platelets coimmunoprecipitation (co-IP) with anti-LDHA or anti–immunoglobulin G (IgG) antibody. The black arrow indicates the location of differential protein. (B) Tandem mass spectrometry assay of eEF2-specific peptide (GGGQIIPTAR) from 95 kD protein gel. (C) Co-IP of DAMI whole-cell lysates with anti-LDHA and anti-eEF2 antibodies. Isotype-matched IgG as negative control (NC). (D) Detection of DNA content (≥4N) in DAMI cells treated with phorbol 12-myristate 13-acetate (PMA; 50 nM), A-484954 (A-484; 10 μM), and DDD107498 succinate (DDD; 1 μM) (n = 3). (E) Functional domains of LDHA and its truncations. Full-length LDHA and its truncated plasmids with Flag-tag (top). Co-IP of coexpression LDHA truncated plasmids and HA-eEF2 plasmid with anti-HA antibody/anti-Flag antibody in HEK-293T (bottom). NC transfected with empty HA/Flag fusion plasmid to control for nonspecific binding. (F) Functional domains of eEF2 and its truncations. Full-length eEF2 and its truncated plasmids with HA-tag. (G) Co-IP of coexpression eEF2-truncated plasmids and Flag-LDHA plasmid with anti-Flag antibody in HEK-293T. (H) Heatmap showing significant differential proteins in LdhA^f/f and LdhA^−/− mice platelets based on quantitative proteomic analysis. Three independent biological replicates were used for cluster analysis, and color represents quantitative ratio. (I) Kyoto Encyclopedia of Genes and Genomes assay of top 10 differential cell pathways in LdhA^f/f and LdhA^−/− mice platelets based on quantitative proteomic analysis. ****P < .0001. PPAR, peroxisome proliferator-activated receptor.