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. 2023 Apr 6;22(5):100544. doi: 10.1016/j.mcpro.2023.100544

Fig. 2.

Fig. 2

Quantitative proteome and phosphoproteome of the rat neurointermediate lobe.A, principal component analysis (PCA) of the NIL proteome and phosphoproteome in control (blue, n = 6) and water-deprived (WD) rats (red; n = 6). B, volcano plot of WD versus control NIL proteome showing 276 upregulated (p-value <0.05, red) and 573 downregulated (p-value <0.05, blue) proteins. C, volcano plot of WD versus control NIL phosphoproteome showing 746 hyperphosphorylation (p-value <0.05, red) and 755 hypophosphorylation (p-value <0.05, blue) events. D, global ΔPs analysis of phosphoproteins between control and WD rats in the NIL. Numbers of hyperphosphorylated (Hyper) and hypophosphorylated (Hypo) peptides are shown. Dotted lines, ΔPs = ±0.4. E, Venn diagram showing 151 proteins in common with changes at the proteome and phosphoproteome level in response to WD. F, phospho raw abundance for S67-p SYN1, S426-p SYN2, and S426-p SYN2 in the NIL according to LC-MS/MS between control (n = 6) and WD (n = 6) rats. Western blotting analysis of S67-p SYN1 (normalized against SYN1), S426-p SYN2 (normalized against SYN2), and S847-p NOS1 (normalized against NOS1) in control (n = 5) and WD NILs (n = 5 for S847-p NOS1 and n = 4 for S67-p SYN1 and S426-p SYN2). G, immunohistochemistry against S67-p SYN1 and arginine vasopressin (AVP), S426-p SYN2 and AVP, and S847-p NOS1 and NOS1 in the pituitary gland of control rats showing the anterior pituitary (AP), intermediate lobe (IL), and posterior pituitary (PP). Images are representative of n = 4. Scale bar represents 75 μm. H, immunohistochemistry against S67-p SYN1, S426-p SYN2, and S847-p NOS1 in the PP of control and WD rats. Images are representative of n = 4. Scale bar represents 25 μm. ΔPs, phosphorylation state change; LC-MS/MS, Nano-LC mass spectrometry; NIL, neurointermediate lobe.