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. 2020 May 4;24(12):6928–6942. doi: 10.1111/jcmm.15351

FIGURE 4.

FIGURE 4

Melatonin attenuates acidosis‐induced synaptic abnormality and tau hyperphosphorylation. A, The differentially expressed proteins related to synaptic‐associated proteins are listed with a ratio (pH6.2/Con), including solute carrier family 17 (sodium‐dependent inorganic phosphate cotransporter), member 6 (SLC17A6), brain acid–soluble protein 1 (BASP1), lymphocyte antigen 6 family member H (Ly6h), thy‐1 membrane glycoprotein (Thy1), eukaryotic elongation factor‐2 kinase (eEF2K), LDL receptor–related protein 6 (LRP6), α‐ketoglutarate‐dependent dioxygenase FTO (FTO), podoplanin (PDPN) and Nckipsd. Red colour indicates the increased proteins (P < 0.05), and green indicates the decreased ones (P < 0.05). To investigate the alterations of synapse‐related proteins, levels of N‐methyl‐D‐aspartate receptor (NR) 2A (NR2A), NR2B, NR1, glutamate receptor (GluR) 1 (GluR1), GluR2, synapsin1, syntaxin1, synaptophysin (Syn), synaptosomal‐associated protein 25 (SNAP25), vesicle‐associated membrane protein 3 (VAMP3) and post‐synaptic density protein 95 (PSD95) were measured by Western blotting (B) and quantitatively analysed (C). D, The differentially expressed proteins related to cytoskeletal system are listed with a ratio (pH6.2/Con), including keratins (Krt1, Krt6a and Krt10), tropomyosins (Tpm1, Tpm3 and Tpm4), neuromodulin (GAP43), SLAIN2 and farnesyl pyrophosphate synthase (FPPS). Red colour indicates the increased proteins (P < 0.05), and green indicates the decreased ones (P < 0.05). Levels of total tau (Tau5), dephosphorylated tau (Tau1) and phosphorylation at Thr231 (pT231), Ser396 (pS396), Ser214 (pS214), Ser404 (pS404) and Thr205 (pT205) of tau were measured by Western blotting (E) and quantitatively analysed (F). To detect the alterations of tau hyperphosphorylation‐related kinases as well as their activity‐related phosphorylation, levels of glycogen synthase kinase 3β (GSK3β), phosphorylated GSK3β at Ser9 (p‐GSK3β), protein kinase B (AKT), phosphorylated AKT at Ser473 (p‐AKT), extracellular‐signal‐regulated kinases (ERK), and phosphorylated ERK at Thr202/Thr204 (p‐ERK), protein phosphatase 2A (PP2A) catalytic subunit (PP2Ac) and its Tyr307‐phosphorylation (p‐PP2Ac) were measured by Western blotting (G) and quantitatively analysed (H). Data were presented as means ± SEM (n = 4/group). *P < 0.05, **P < 0.01, ***P < 0.001 vs vs Con, # P < 0.05, ## P < 0.01, ### P < 0.001 vs Veh