Fig 1.
Normal pancreas (NP) and caerulein-induced acute pancreatitis (AP).
Representative images of H&E stained histology slides of A) NP with intact pancreas architecture and B) AP showing marked oedema, inflammatory cell infiltration and acinar cell necrosis. Mean serum amylase levels in (C) NP and (D) AP in each experiment consisting of 16 individuals.
Fig 2.
Preparation of a plasma membrane enriched fraction.
Coomassie-stained SDS-PAGE gel of (A) NP and (B) AP samples obtained during homogenisation and fractionation by sequential steps of centrifugation. Nu = nuclear pellet; S1 = post-nuclear supernatant; Mt = mitochondrial pellet; S2 = post-mitochondrial supernatant; C = cytosol (post-microsomal supernatant); W = wash of the microsomal pellet; Mc = microsomal pellet. Coomassie-stained SDS-PAGE and western blot analysis of 10 fractions (F1-F11) from the microsomal pellet after flotation on a sucrose gradient (0.25–2 M) in (C) NP and (D) AP. Fractions are ordered depending on their equilibrium density from light (left) to heavy (right). The enrichment of plasma membrane was assessed by western blot using an antibody against caveolin-1, which is a specific plasma membrane marker. Full Western blots in S1 Fig.
Fig 3.
Heat map depicting the variation across the biological and technical replicates in extracellular pancreas HBPs.
The rows represent the various biological replicates in normal pancreas (NP) and acute pancreatitis (AP), while the columns represent proteins. Red represents over expression and green represents under expression. Biological replicate number is denoted as "BioRep" and technical replicate number as "TechRep". Hierarchical clustering was performed on both column data, to cluster the changes in protein expression, and row data, which displays the variation between samples. The stability of the instrument platform is shown in that the lowest branch of the sample variation dendrogram correctly represents the technical replicates of each sample. The next level of the dendrogram correctly separates the biological condition of the sample indicating repeatable protein expression differences between the biological conditions. A higher degree of variability is observed in the AP samples presumably reflecting the systemic effects of AP.
Table 1.
Top 20 extracellular pancreas HBPs overexpressed in AP.
The upregulated HBPs were filtered depending on the maximum fold change values. An adjusted threshold p value of less than 0.001 following the Bonferroni correction was used to identify the top HBPs to be validated as potential biomarkers.
Table 2.
Top 20 extracellular pancreas HBPs underexpressed in AP.
The downregulated HBPs were filtered depending on the maximum fold change values. An adjusted threshold p value of less than 0.001 following the Bonferroni correction was used to identify the top HBPs to be validated as potential biomarkers.
Fig 4.
The heparin-binding putative protein interactome in normal pancreas (NP) constructed using STRING 10.5.
Nodes or HBPs are connected by protein-protein interactions known as ‘edges’.
Fig 5.
The heparin-binding putative protein interactome in acute pancreatitis (AP) constructed using STRING 10.5.
Nodes or HBPs are connected by protein-protein interactions known as ‘edges’.
Table 3.
Top 20 canonical pathways enriched to extracellular pancreas HBPs in NP using ingenuity pathways analysis.
The significance of the association between the datasets and the canonical pathway was measured by calculating the p-value using Fisher’s exact test to determine the probability of the association between the HBPs in the dataset and the canonical pathway.
Table 4.
Top 20 canonical pathways enriched to extracellular pancreas HBPs in experimental AP using ingenuity pathways analysis.
The significance of the association between the datasets and the canonical pathway was measured by calculating the p-value using Fisher’s exact test to determine the probability of the association between the HBPs in the dataset and the canonical pathway.
Fig 6.
Heat map depicting the variation across the biological and technical replicates in plasma HBPs.
The rows represent the various biological replicates from plasma in health (NP) and acute pancreatitis (AP), while the columns represent proteins. Red represents over expression and green represents under expression. Biological replicate number is denoted as "BioRep" and technical replicate number as "TechRep". Hierarchical clustering was performed on both column data, to cluster the changes in protein expression, and row data, which displays the variation between samples.
Table 5.
Top 20 plasma HBPs overexpressed in AP.
The upregulated HBPs were filtered depending on the maximum fold change values.