Fig. 2.

The seminal fluid proteome responds differentially to aging in U versus F males. (A) Heatmap of the abundance of the 117 Sfps detected in accessory gland and ejaculatory duct samples (n = 4 replicate experiments per group). The abundance of 40 out of 117 Sfps exhibit a significant differential response to age and mating after multiple test corrections. The annotation classification of each Sfp is indicated. (B) Line plots showing the change in standardized Sfp abundance with age. The average change in Sfp abundance for U and F males is depicted with lines marked U and F, respectively (age and mating interaction: L ratio²₂ = 163.856; P < 0.0001). (C) Principal component analyses of the seminal fluid proteome in male reproductive tissues (age and mating interaction: L ratio²₂ = 34.949; P < 0.0001). (D) Heatmap of the abundance of 117 seminal fluid proteins transferred to females during mating. None of the individual 117 Sfps exhibited a significant interaction between age and mating group after multiple test corrections. Two ejaculatory duct-specific Sfps were transferred in significantly higher quantities in response to age, independent of mating activity (CG17242 and CG5162), and 10 Sfps were transferred in significantly higher quantities in response to frequent mating independent of age (Acp26Aa, CG10587, CG17472, CG3097, CG34002, Est-6, NLaz, Regucalcin, Sfp24F, and Sfp65A). The annotation classification of each Sfp is indicated. (E) Line plots showing the standardized abundance of Sfps transferred with age. The average change in Sfp abundance for U and F males is depicted with lines marked U and F, respectively (age and mating interaction: L ratio²₂ = 130.595; P < 0.0001). (F) Principal component analyses of the seminal fluid proteome transferred to females (age and mating interaction: L ratio²₂ = 11.485; P = 0.003). Differences at P < 0.05 within mating groups and age categories are represented as different letters.