Fig 1: Immunoblotting analysis. (A) 2-DE gel images of six protein spots of interest denoted by an arrow in each panel. Each panel shows an expanded 2-D gel view and ranked from the left to the right: the left (CL) and the right hippocampus (CR) of the control group; the left (PTZL) and the right hippocampus (PTZR) of the PTZ group; the left (ElecL) and the right hippocampus (ElecR) of the electrical kindling group; and the left (PiL) and the right hippocampus (PiR) of the pilocarpine group. (B) Western blot analysis validates the differential expression of ADPRC1, Calreticulin, SNAP 25, UCH-L1, LPAR3 and transgelin 3 in the control and the epileptic groups. Beta-actin was used for normalization. (C) The intensity of bands was quantified by ImagJ software. The data were expressed as mean ± S.E.M. The letters above the columns (a-h) indicate significant differences (P < 0.05) using the Duncan’s multiple range test so, data that showed with the same letter don’t have significant differences. (D) Representative photographs of immune-histochemical analysis of the expression levels of the ADPRC1, the calreticulin, the SNAP 25 and the UCH-L1 in control and epileptic groups.
Fig 2: The expression levels of TREM2 (triggering receptor expressed by myeloid cells 2), DAP12 (DNAX-activation protein 12), IBA1, cleaved caspase 3, SNAP25 and PSD95 by disease groups. The expression levels of microglial proteins TREM2, DAP12 and IBA1, apoptotic marker active caspase 3, and synaptic proteins SANP25 and PSD95 were analyzed in the brain samples using protein-specific antibodies by our standard immunoblotting procedure. The intensities of the specific band in each sample were normalized by ß-actin followed by statistical analysis by disease groups: nondemented controls (ND); possible AD (PossAD); and Alzheimer's disease (AD). Significant changes (P < 0.05) were detected between AD and ND groups in TREM2 (A), DAP12 (B), IBA1 (C), and active caspase 3 (D), SNAP25 (E) and PSD 95 (F). However, the significant differences between AD and PossAD groups were only detected in TREM2, DAP12 and IBA1. Expression levels of the apoptotic marker active caspase 3, presynaptic protein SNAP25 and post-synaptic protein PSD95 were not significantly different between PossAD and ND groups. There were no significant differences between ND and PossAD in all of the markers. The horizontal lines in the dot plots represent the values of mean and standard errors. Representative immunoblot images for the proteins TREM2, DAP12, IBA1, PSD95, SNAP25 and cleaved caspase 3 were compiled in G. The disease groups for the samples in each lane were shown above gel images: AD for Alzheimer's disease; PA for possible Alzheimer's disease, ND for nondemented normal controls. The blank lane was the lane that molecular weight ladders were loaded. The molecular weights corresponding to each protein were indicated at the right side of the gel images. The brain TREM2 has molecular weights of 35–40 kDa.
Supplier Page from Abcam for Anti-SNAP25 antibody [EP3274]