The expression of TMEM106A, TMEM106B, TMEM106C, and PGRN mRNAs is thus universal in human neural cell lines. Reduced expression of TMEM106B mRNA in Alzheimers disease brains We next analyzed by qPCR the levels of TMEM106B, PGRN, and G3PDH mRNAs in frozen human brain tissues derived from four NC cases, six ALS cases, four PD cases, and seven AD cases presented in Table 1. Before starting despite this, we investigated the p. T185S genotype of rs3173615 in the human TMEM106B gene, on which the T185 iso form acts as a risk factor, while the S185 isoform serves NFH mRNA expression Inhibitors,Modulators,Libraries levels, the T185 Inhibitors,Modulators,Libraries S185 heterozygote consisted of 14 cases, and the S185 S185 homozygote consisted of three cases, consistent with the genotyping data of HapMap JPT. The frequency of T185 and S185 isoforms Inhibitors,Modulators,Libraries was thus not significantly different between AD and non AD groups.
By qPCR, AD cases showed significantly reduced mRNA levels Inhibitors,Modulators,Libraries of TMEM106B, when compared with those in non AD cases. In contrast, AD cases showed significantly elevated mRNA levels of PGRN, with some variations among the cases, when compared with the levels in non AD cases. Not ably, a significant negative correlation was found between TMEM106B and PGRN mRNA expression levels. Furthermore, AD cases showed significantly reduced mRNA levels of NFH and elevated mRNA levels of GFAP and NEUN, when compared with the levels in non AD cases. Importantly, a significant positive correlation was found between TMEM106B and coefficient 0. 496, P 0. 0221 . Moreover, we studied by qPCR the levels of TMEM106A and TMEM106C mRNAs in AD and non AD brains.
Both were markedly elevated Inhibitors,Modulators,Libraries in AD brains, compared with the levels in non AD brains. The expression of TMEM106B paralo gues was uniquely regulated in the opposite direction to the expression levels of TMEM106B. Characterization of the specificity of anti TMEM106B antibody The specificity of anti human TMEM106 antibody was veri fied by western blot of recombinant human TMEM106A, TMEM106B, and TMEM106C proteins tagged with Xpress expressed http://www.selleckchem.com/products/crenolanib-cp-868596.html in HeLa cells. The A303 439A anti TMEM106B antibody recognized 45 kDa monomeric and 120 kDa oligo meric forms of TMEM106B tagged with Xpress, whereas it did react either with TMEM106A or with TMEM106C, validating the specificity of the A303 439A antibody. In contrast, both bs 11694R and 20995 1 AP anti TMEM106B antibodies did not specifically react with the Xpress tagged human TMEM106B protein. We therefore se lected A303 439A for western blot and immunohistochem istry analysis in the present study. This antibody specifically reacted with a major 31 kDa protein endogenously expressed in human brain tissues and IMR 32 neuroblast oma cells.