Journal: Aging Cell

Article Title: Long‐term exposure to excessive norepinephrine in the brain induces tau aggregation, neuronal death, and cognitive deficits in early tau transgenic mice

doi: 10.1111/acel.14420

Figure Lengend Snippet: RBX facilitates the activities of tau kinases, PKA and GSK3β. (a, b) Representative western blot images. (c, d) Quantification of specific antibodies against pPKA, PKA, pGSK3β (S9), pGSK3β (Y216), or GSK3β in the hippocampus following treatments in the ADLP Tau mice. One dot in the bar graphs represents each mouse (c, d). All data represent mean ± SEM. Significance was determined by unpaired t test and denoted by * p < 0.05, ** p < 0.01, *** p < 0.001, or **** p < 0.0001.

Article Snippet: The following day, the cells were transfected with plasmids encoding GSK3β WT (Addgene, #14753), GSK3β S9A (Addgene, #14754), or GSK3β K85A (Addgene, #14755) using Lipofectamine LTX (Thermo Fisher Scientific), following the manufacturer's protocol.

Techniques: Western Blot

Journal: Aging Cell

Article Title: Long‐term exposure to excessive norepinephrine in the brain induces tau aggregation, neuronal death, and cognitive deficits in early tau transgenic mice

doi: 10.1111/acel.14420

Figure Lengend Snippet: Human brain organoids exposed to the elevated NE facilitate the activities of PKA and GSK3β and aggravate tau aggregation. (a) Schematic representation of the experimental design for generating human brain organoids. (b–f) Representative western blot images and quantification of specific antibodies against 6E10, AT8, Tau13, pPKA, PKA, pGSK3β (Y216), or GSK3β in the human brain organoids exposed to various levels of NE. One dot in the bar graphs represents each organoid (b–f). All data represent mean ± SEM. Significance was determined by one‐way ANOVA and denoted by * p < 0.05, ** p < 0.01, *** p < 0.001, or **** p < 0.0001.

Article Snippet: The following day, the cells were transfected with plasmids encoding GSK3β WT (Addgene, #14753), GSK3β S9A (Addgene, #14754), or GSK3β K85A (Addgene, #14755) using Lipofectamine LTX (Thermo Fisher Scientific), following the manufacturer's protocol.

Techniques: Western Blot

Journal: Molecular Biology of the Cell

Article Title: Amyloid-β oligomers induce tau-independent disruption of BDNF axonal transport via calcineurin activation in cultured hippocampal neurons

doi: 10.1091/mbc.E12-12-0858

Figure Lengend Snippet: Inhibition of protein phosphatase-1 and glycogen synthase kinase 3β prevents AβO-induced transport defects. (A) Expression of myc-tagged I-2 and BDNF-mRFP in an AβO-treated tau −/− neuron (from left to right). Overlay of BDNF-mRFP and AβO images shows binding of AβOs exclusively to dendrites. Retrospective immunocytochemistry was used to confirm I-2 expression and AβO binding. Arrows indicate axon; arrowheads indicate dendrites. Scale bar, 50 μm. (B) Effects of PP1 and GSK3β inhibition on BDNF flux in AβO-treated tau +/+ and tau −/− neurons. Bidirectional flux is markedly reduced by AβOs, and transport defects are prevented by expression of I-2 for 24 h, pretreatment with 5 μM inhibitor VIII for 30 min, and expression of kinase-dead GSK3β (K85A) for 24 h. Flux is similarly impaired by expression of kinase-active GSK3β (S9A). (C) Representative kymographs comparing BDNF transport in control, AβO-treated, and inhibitor-treated tau +/+ and tau −/− neurons. A minimum of 15 cells from three different cultures were analyzed per condition; *** p < 0.001 relative to controls and +++ p < 0.001 relative to AβO-treated cells. Complete statistical evaluation is presented in Supplemental Tables S3 and S4.

Article Snippet: Experiments assessing a role for GSK3β and PP1 used the following plasmids and were transfected as described: pcDNA3 HA-GSK3β S9A and pcDNA3 HA-GSK3β K85A (gifts from Jim Woodgett, University of Toronto; plasmids 14754 and 14755; Addgene, Cambridge, MA) and I-2/pCS2 (plasmid 16317; Addgene).

Techniques: Inhibition, Expressing, Binding Assay, Immunocytochemistry, Control