Journal: Frontiers in Neurology

Article Title: Biochemical Distribution of Tau Protein in Synaptosomal Fraction of Transgenic Mice Expressing Human P301L Tau

doi: 10.3389/fneur.2014.00026

Figure Lengend Snippet: Soluble and sarkosyl-insoluble tau in JNPL3 male mice . (A) Western blots of TBS-soluble tau in mouse cerebral cortices. Equal volumes of TBS-soluble fraction derived from 0.2 mg wet weight of brain from eight male JNPL3 and two male non-tg mice were separated by SDS-PAGE, blotted, and then probed with E1, Tau5, MS06, and GAPDH antibodies. (B) Western blot of sarkosyl-insoluble fractions in mouse cerebral cortices. Samples derived from 20 mg wet weight from male JNPL3 and non-tg mice, 5 mg wet weight from female JNPL3 cortex (ctx), and 2.5 mg wet weight from female JNPL3 spinal cord (SPc) were separated by SDS-PAGE, blotted, and then probed with E1 antibody.

Article Snippet: The samples were separated by gel electrophoresis on 10 or 5–20% gradient SDS-PAGE gels (Wako Pure Chemical Industries, Osaka, Japan), and transferred to nitrocellulose membranes (Schleicher & Schuell BioScience, Dassel, Germany).

Techniques: Western Blot, Derivative Assay, SDS Page

Journal: Frontiers in Neurology

Article Title: Biochemical Distribution of Tau Protein in Synaptosomal Fraction of Transgenic Mice Expressing Human P301L Tau

doi: 10.3389/fneur.2014.00026

Figure Lengend Snippet: Subcellular fractionation of mouse cerebral cortex . (A) Schematic representation of the subcellular fractionation steps. P1, nuclear pellet and debris; P2, crude synaptosomal fraction; P3, light membranes; S3, cytosolic fraction; LP1, synaptosomal membrane fraction; LP2, synaptic vesicle-enriched fraction; LS2, soluble synaptosomal fraction. (B) Western blots of JNPL3 and non-tg male mouse cerebral cortex subcellular fractions. P1, P2, P3, or S3 fraction derived from 0.13 mg wet weight of tissue and LP1, LP2, or LS2 fraction derived from 0.5 mg wet weight of tissue were loaded on SDS-PAGE. Blots were probed with PSD-95, synaptotagmin, GAP-43, Tau5, E1, β-tubulin, and β-actin antibodies. (C) Proportions of protein levels in fractions (P1, P2, P3, and S3) of tau (Tau5) and β-tubulin (β-tub) from JNPL3 ( n = 5) and non-tg ( n = 5) mice are shown. Intensities of tau (49–65 kDa) and β-tubulin (50 kDa) were measured by Bio-Imaging Analyzer System. Ratios were indicated by percent of total (P1 + P2 + P3 + S3). Results are expressed as mean ± SEM. (D) Proportions of protein levels in synaptosomal fractions (LP1, LP2, and LS2) of tau (Tau5) and β-tubulin (β-tub) from JNPL3 ( n = 5) and non-tg ( n = 5) mice were indicated. Intensities of tau (49–65 kDa) and β-tubulin (50 kDa) were measured by Bio-Imaging Analyzer System. Ratios were indicated by percent of total (LP1 + LP2 + LS2). Results are expressed as mean ± SEM.

Article Snippet: The samples were separated by gel electrophoresis on 10 or 5–20% gradient SDS-PAGE gels (Wako Pure Chemical Industries, Osaka, Japan), and transferred to nitrocellulose membranes (Schleicher & Schuell BioScience, Dassel, Germany).

Techniques: Fractionation, Western Blot, Derivative Assay, SDS Page, Imaging

Journal: Frontiers in Neurology

Article Title: Biochemical Distribution of Tau Protein in Synaptosomal Fraction of Transgenic Mice Expressing Human P301L Tau

doi: 10.3389/fneur.2014.00026

Figure Lengend Snippet: Quantitative western blot analysis of tau protein . (A) Tau band patterns in cytosolic (S3) and synaptosomal membrane (LP1) fractions from four JNPL3 and four non-tg mouse cerebral cortices. Equal volumes of fractions derived from 0.25 mg wet weight of brain were separated by SDS-PAGE, blotted, and then probed with Tau5, MS06, Tau1, pS199, pT231, and pS396 antibodies. (B,C) The relative ratio of tau protein between S3 and LP1 fractions from JNPL3 (B) and non-tg (C) mice was measured ( n = 4 each). Results are expressed as mean ± SEM. The mean value of tau protein in S3 fraction from JNPL3 mice was normalized to one.

Article Snippet: The samples were separated by gel electrophoresis on 10 or 5–20% gradient SDS-PAGE gels (Wako Pure Chemical Industries, Osaka, Japan), and transferred to nitrocellulose membranes (Schleicher & Schuell BioScience, Dassel, Germany).

Techniques: Western Blot, Derivative Assay, SDS Page