Figure 6

Label-free proteomics analysis of intestinal mucus from 3 animals per age group. A Heatmap of sample correlations after hierarchical clustering using Spearman correlation coefficients between protein expression values of all pairs of samples. B Venn diagram showing the unique and shared proteins between 3-day old pig and 3-week-old pig determined by the label-free analysis. C A volcano plot was generated for every protein identified using a log₂ fold change of 3-days/3weeks on the X-axis, and -log₁₀ adjusted p-value on the Y-axis. D Mean iBAQ values ± SD of identified mucins from the mucus of three animals per age group are plotted as scatter graphs with bars. Statistically significant differences, determined by two-way ANOVA (followed by Šídák’s multiple comparison post hoc test), are represented as *(p < 0.05) or ***(p < 0.001). E Heatmap showing the differences of z-scored values of the abundance of selected proteins from six categories (mucins, coronavirus receptors, sugar transferases, antimicrobial peptides, cell integrity proteins, and digestive enzymes) between each sample across 6 replicates (3 from each age-group). F The hierarchical clustering of selected proteins from six categories and 6 replicates across two age-groups using Euclidian distance is shown in the heat map. For both E and F, the heat map color variations shows the maximum and minimum protein abundance between each replicate of the two age groups (row min/row max).