Table 3 Functional Annotations analysis of DEPs by DAVIDS analysis in low, medium and high ROS group

Functional categories related to reproduction/spermatogenesis

Single fertilization (4), sexual reproduction (8)

Sexual reproduction (7), gamete generation (96), reproductive process in multicellular organism (6)

Sexual reproduction (11), multicellular organism reproduction (10), gamete generation (9), spermatogenesis (8), spermatid development/differentiation(3), germ cell development (3)

Enriched functional categories

Cell cortex (5), actin binding (5), myofibril (3), protease (4), peptidase activity (4), metal ion binding (12), protein targeting (3), IC protein transport and localization (3), cell cycle phase (3), nucleotide binding (5), regulation of transcription (3); lysosome (7), influence lipid concentrations and risk of CAD (4), plasma lipoprotein particle remodeling (3), heparin binding (5), protein-lipid complex (3), glucose metabolic process (5), cytoskeleton organization (6), sterol transport and homeostasis (3), EGF-like region (5), cellular response to stress (3), negative regulation of apoptosis (6), cell migration (5), cell cycle (3)

Basement membrane (6), EC matrix (6), ECM-receptor interaction (5), EGF domain (5), cell motility and localization (6), cytoplasmic vesicle (6), regulation of neurological system process (3), motor protein (3), calmodulin binding (3), metal ion binding (16) cellular ion homeostasis (3), cell morphogenesis (3), protein targeting and localization (3), synaptic transmission (3), ATP binding (5); Dynein (6), microtubule associated complex (6), motor activity (6), negative regulation of cellular metabolic process (5), Ca binding region (4), cytoskeleton organization (5), gamete generation (6), reproductive process in multicellular organism (6), nucleotide biosynthetic process (4), nucleotide binding (12), metalloendopeptidase activity (3), glycosaminoglycan binding (3), phospholipid metabolic process (3), regulation of apoptosis (6), cytoskeleton (11), metal ion binding (12)

Oxidoreductase (12), Protease (11), cofactor binding (8), metallopeptidase activity (7), metalloprotease (7), zinc ion binding (13), cytoskeleton (14), cell junction (6), regulation of apoptosis (7), intracellular protein transport and localization (5), proteasome complex (3), vasculature development (4), focal adhesion (4); nucleotide binding (24), sexual reproduction (11), multicellular organism reproduction (10), gamete generation (9), spermatogenesis (8), spermatid development/differentiation(3), germ cell development (3), glutathione S-transferase (3), peroxisome (4), fatty acid oxidation (3), lipid modification (3), glycerolipid metabolic process (4), protein targeting (3), metal ion binding (13), apoptosis (3)

Majority of proteins associated with functions

Polymorphism (29), phosphoprotein (22), acetylation (15), glycoprotein (13), Golgi (9); polymorphism (62), signal peptide (38), glycoprotein (36), disulfide bond (34), extracellular (30), acetylation (26), secreted (23), oxidoreductase (11), lipid binding (11)

Phosphoprotein (26), glycoprotein (20), signal peptide (18), disulfide bond (15), acetylation (13), cytoplasm (13), Golgi (10), EC region (11), secreted (10), Ca ion binding (8), cell adhesion (7); Polymorphism (50), Signal peptide (18), EC (17), secreted (15), mitochondrion (13), nucleotide binding (15), cell fraction (10), cytoskeleton (9), sexual reproduction (7)

Polymorphism (57), phosphoprotein (41), ion binding (34), glycoprotein (27), cytoplasm (26), signal peptide (22), cell membrane (16), proteolysis (14), hydrolase (15), oxidoreductase (12), defense response (9), metalloprotease (7), protein transport and localization (8), exopeptidase activity (6), adherens junction (6), cell migration (6); purine nucleotide binding (24), ribonucleotide binding (22), sexual reproduction (11), gamete generation (9), spermatogenesis (8), mitochondrion (15), reproductive cellular process (5), protein folding (5), spermatid development (3)

Cellular distribution

Cell cortex (5), Golgi (9), cytoskeleton (9), cytosol (8), ER (6), organelle envelope (5), vesicle (5), sarcomere (3); extracellular region (30), cytosol (13), lysosome (7), vesicle (11), secretory granule (4), protein-lipid complex (3), cell surface (6), golgi (10)

Golgi (10), EC region (9), organelle membrane (9), ER (8), proteinaceous ECM (6), cytoplasmic vesicle (6), intrinsic to plasma membrane (8); dynein complex (6), mitochondrion (13), EC region (17), microtubule cytoskeleton (8), cell projection (8), cytoskeleton (11), cilium (4)

Actin cytoskeleton (10), cell cortex (7), cortical cytoskeleton (5), cell-cell adherens junction (4), vesicle (11), Golgi (12), EC region (19), mitochondrial lumen (4); mitochondrion (15), cytosol (15), dynein complex (3), microbody (4), peroxisome (4), cell fraction (11), secretory granule (4), organelle envelope (7), pore complex (3)

Activated processes/functions

Intracellular transport (7), protein localization (7), cytoskeletal organization (5), regulation of protein complex assembly (3), microtubule based process (4), calmodulin binding (4), motor activity (4), actin binding (5), serine hydrolase activity (3), phospholipid binding (3)

Cell motility and localization (6), cellular iron ion hemostasis (3), cell migration (5), cell motion (6), cell adhesion (7), integrin-mediated signaling pathway (3), macromolecular complex assembly (6), response to wounding (5), regulation of neurological system process (3), Ca ion binding (8), peptidase activity (5), carbohydrate binding (4), microfilament motor activity (2), structural molecule activity (5)

Actin filament based process (9), oxidation reduction (12), integrin mediated signaling pathway (5), cytoskeleton organization (9), proteolysis (14), cell motion (9), defense response (9), intracellular transport (8), cell-cell adhesion (5), oxygen and ROS metabolic process (3), actin binding (10), exo- or endo-peptidase activity (6), cytoskeletal protein binding (11), calcium ion binding (11), cofactor binding (8), peptidase activity (11), antioxidant activity (3)

Downregulated processes/functions

Glycerolipid metabolic process (6), lipid hemostasis (4), oxidation reduction (11), single fertilization (4), protein-lipid complex remodeling (3), sexual reproduction (8), protein folding (5), sterol transport (3), response to oxidative stress (5), lipid binding (11), carbohydrate binding (8), glycosaminoglycan binding (5), oxidoreductase activity (3), antioxidase activity (3)

Actin filament organization (4), protein folding (5), negative regulation of cellular metabolic process (5), sexual reproduction (7), protein tetramerization (3), oxidation reduction (8), gamete generation (6), cell redox homeostasis (3), protein oligomerization (4), generation of precursor metabolites and energy (5); motor activity (6), enzyme inhibitor activity (6), proteasome regulator activity 92), protein binding (7), nucleoside binding (12), nucleotide binding (15), protein homodimerization activity (5)

Sexual reproduction (11), reproductive process in a multicellular organism (10), gamete generation (9), spermatogenesis (8), protein folding 95), glycerophospholipid metabolic process (4), spermatid development/differentiation (3); purine nucleotide binding (24), ATP binding (18), microtubule motor activity (3), sterol transporter activity (2), cAMP dependent protein kinase regulator activity (2), glutathione transferase activity (2)

Activated pathways

Ether lipid metabolism (2), Synaptic proteins at the synaptic junction (2);

ECM-receptor interaction (5), Integrin cell surface interactions (4)

Leukocyte transendothelial migration (5), focal adhesion (8), ECM-receptor interaction (4), Adherens junction (4), Hemostasis (7), Apoptosis (5), Metabolism of lipids and lipoproteins (5), metabolism of amino acids (5), Wntsignaling (3), Integrin signaling pathway (3), Tight junction (4), Proteasome (3)

Downregulated pathways

Gluconeogenesis (3), metabolism of carbohydrates (4), hemostasis (5)

Fructose and mannose metabolism (3), apoptosis (4), metabolism of amino acids (4)

Glycolysis/gluconeogenesis (3), adipocytokine signaling pathway (3), PPAR signaling pathway (3), metabolism of lipids and lipoproteins (5), Integration of energy metabolism (5), mechanism of protein import into the nucleus (2)

Processes/Functions unique to each ROS group

Regulation of cellular protein metabolic process (2), endocytosis (1), PTM (1)

Organelle fusion (2), protein homodimerization activity (2), anti-apoptosis (2), ribosomal subunit (2), cytosol (3), membrane (4), polymorphism (5)

Disulfide bond (7), signal peptide (6), alternative splicing (8), metal ion binding (7), glycoprotein (8), secreted (4), endocytosis (2), aging (2), plasma membrane (5), calcium ion binding (5)

Processes/Functions unique to Control group

MAPK signaling pathway (2), Regulation of actin cytoskeleton (2), metabolism of vitamins and cofactors (2)

Tissue morphogenesis (3), tube development (3), embryonic morphogenesis (3), leucine rich repeat (3), extracellular region (5), glycoprotein (5), membrane (3), urogenital system development (2)

Transmembrane (3), alternative splicing (3), transport (4), metabolism of vitamins and cofactors (1)