Figure 5.

Proteomic analysis of insulin-stimulated adipose tissue from postpartum (PP) dairy cows supplemented with ALA. Dairy cows were divided into two nutritional regiment groups from 21 to 60 days PP; (i) CTL—encapsulated saturated fat, (ii) ALA—flaxseed supplement providing α-linolenic acid (ALA). n = 5 per treatment. (A) Work flow for the proteomic analysis; proteins extracted from adipose were digested and peptides were identified for their differential expression. Bioinformatic analysis and network analysis were performed to identify the enriched pathways. Western blot analysis was used for validation. (B) Principal component analysis (PCA) of the AT proteome; CTL samples are denoted in red circles, whereas ALA samples are denoted in blue triangles. (C) Heat map analysis of AT proteome: low peptide intensity is denoted in green, whereas high intensity is denoted in purple. Each cow in the study is numbered and represented in columns. (D) Volcano plot analysis of the AT proteome; the P value (< 0.05) is represented on the Y-axis and FDR (± 1.5) is represented on the X-axis. Each dot represents one protein: red denotes more, whereas blue denotes fewer proteins in AT. (E) Correlation of CTL vs ALA cows generated by the IDEP9.5 server where c1, c2, c3, c4, and c5 represent CTL; s1, s2, s3, s4, and s5 represent ALA-supplemented cows. (F) GO analysis of AT proteome supplemented with ALA including the biological process category, the molecular function category, and the cell component category. Image was generated using www.Biorender.com.