Journal: Brain pathology (Zurich, Switzerland)

Article Title: OVERLAPPING BUT DISTINCT TDP-43 AND TAU PATHOLOGIC PATTERNS IN AGED HIPPOCAMPI

doi: 10.1111/bpa.12505

Figure Lengend Snippet: This is the result of analyses of a subsample of 6 cases with 2 slides/case analyzed, comprising 14,200 dentate granule neurons from CARTS cases and 11,952 cells from AD cases. Panel A shows the overall percentage of immunopositive cells. Note that approximately 1% of cells are positive for PHF-1 and TDP-43, with trend for more PHF-1 immunoreactive cells in the dentate granule cell layer from AD cases. Panel B shows the percentage of cells that are colocalized among those that are immunopositive. Panel C shows that in both AD and CARTS, a higher percentage (~25%) of cells that were TDP-43 immunopositive were also PHF-1 immunopositive, in comparison to the colocalization among the PHF-1 immunopositive cells in terms of p-TDP-43 immunopositivity (4%) **p=0.007.

Article Snippet: By contrast, the arrowhead in (B) shows a PHF-1 positive cell that is colocalized with P-TDP-43, as shown at a higher magnification in (D) . fig ft0 fig mode=article f1 fig/graphic|fig/alternatives/graphic mode="anchored" m1 Open in a separate window caption a7 caption a8 Cell counting and colocalization method used Imaris 3D microscopic data analysis software on confocal Z-stack images (A) PHF-1 was thresholded to exclude pixels detected in the no PHF-1 primary antibody control samples. (B) P-TDP-43 was similarly thresholded to exclude pixels detected in the no P-TDP-43 primary antibody control samples. (C) Using the Imaris Coloc function a new channel was generated that corresponded to the pixels (after thresholding) that were in present in both the PHF-1 and P-TDP-43 channel.

Techniques: Comparison

Journal: Journal of Cell Science

Article Title: Myosin VI mediates the movement of NHE3 down the microvillus in intestinal epithelial cells

doi: 10.1242/jcs.149930

Figure Lengend Snippet: A pool of myosin VI localizes on the microvillus of intestinal epithelial cells. (A) Immunofluorescence (confocal microscopic) detection of myosin VI expression and localization in Caco-2/Bbe cells and mouse ileum. In Caco-2/Bbe cells, myosin VI is present throughout the brush border, including in the microvillus and in a vesicular pattern in the terminal web and other intracellular compartments (a1–a4). In mouse ileum, myosin VI is present to a lesser extent on the microvillus and in the terminal web (b1–b4). As a negative control, anti-myosin VI antibody was used to show there is no myosin VI staining in ileum from myosin VI KO mice (d) and myosin VI KD Caco-2/Bbe cells (f). (c) wild-type (WT) control; (e) empty-vector-infected control. Scale bar: 20 µm. (B) Analysis of the overlap of myosin VI and protein markers of microvillus by MetaMorph colocalization software. Please note this software shows presence of two proteins in the same intracellular compartment but does not establish physical association. In Caco-2/Bbe cells, ∼46% of myosin VI overlaps with villin, and in mouse ileum ∼9% of myosin VI overlaps with aquaporin 7. For both Caco-2/Bbe cells and mouse ileum, results are mean±s.e.m. of three separate experiments with five or six images analyzed in each experiment (the total number of images analyzed was 16).

Article Snippet: Overlap of myosin VI with the microvillus marker aquaporin 7 was determined with MetaMorph software colocalization analysis.

Techniques: Immunofluorescence, Expressing, Negative Control, Staining, Plasmid Preparation, Infection, Software

Journal: Neurobiology of aging

Article Title: Dystrophic microglia are associated with neurodegenerative disease and not healthy aging in the human brain

doi: 10.1016/j.neurobiolaging.2020.12.003

Figure Lengend Snippet: A quantitative cellular localization of FTL found colocalization FTL with microglia. FTL did not appear to be staining all microglia; therefore, we sought to see if there was any association of FTL with the different microglia morphologies we recently characterized. (A) Shows an example of the IBA1+FTL+ staining. (B) Shows the HALO generated markup, where green is IBA1, red is FTL, and yellow is the area where the two proteins are colocalized. (C) Shows that there was greater IBA1 staining intensity in the hypertrophic microglia compared to the two different morphologies for the IBA1 markup area (p<0.0001, Tukey Test). (D) A high degree of IBA1+FTL+ area of colocalization in the dystrophic microglia compared to the ramified or hypertrophic morphologies (p<0.0001, Tukey Test). Circles are individual microglia, from 3 different aged brains.) The scale bar is 20 μm.

Article Snippet: We then used the area colocalization FL algorithm (Halo software, version 2.3; Indica labs) on the ROIs to determine the area of colocalization of IBA1 and FTL for the three microglia morphologies.

Techniques: Staining, Generated