Journal: Nature neuroscience

Article Title: Therapeutically viable generation of neurons with antisense oligonucleotide suppression of PTB.

doi: 10.1038/s41593-021-00864-y

Figure Lengend Snippet: Fig. 3 | Transition of GFAP:tdTomato+ cells into new neurons results in no depletion in the total GFAP cell number. a, Images of newly induced neurons (iNeurons) expressing NeuN (green) and tdTomato (red) (NeuN visualized by IF and tdTomato by direct fluorescence); blue, DAPI staining for DNA; white, GFAP visualized by IF; the experiment was reproduced four times, independently, with similar results. b, Total number of tdTomato+NeuN− cells (left) or tdTomato+NeuN+ cells (right), either with or without GFAP labeling, per dentate gyrus, 2 months after ICV injection of PTB-ASO. Data are presented as mean ± s.e.m. (left: tdTomato/NeuN: 176.8 ± 34.92; tdTomato/NeuN/GFAP mean: 167 ± 34.32; right: tdTomato/NeuN mean: 28.88 ± 4.87; tdTomato/NeuN/GFAP mean: 0.87 ± 0.12; n = 4 biological repeats; two-tailed t-test **P = 0.0012). c, Representative images of human organoids 1 month after PTB-ASO or control-ASO application in the growth media GFAP (green) and Nestin (red) (both visualized by IF); blue, DAPI staining for DNA; the experiment was reproduced three times, independently, with similar results. d, Representative images of a dentate gyrus taken 2 months after ICV injection of control or PTB-ASO2 into 3-month-old and 1-year-old mice; white, GFAP visualized by IF; the experiment was reproduced three times for control and aged mice and four times for 5-month-old mice, independently, with similar results. e, Total GFAP-positive cells per dentate gyrus area of 5-month- and 1.2-year-old PTB-ASO- or control-injected mice. Data are presented as mean ± s.e.m. (control mean: 1,074 ± 83.55; 5 months PTB-ASO mean: 1,006 ± 82.45; 1.2 year-old PTB-ASO mean: 1,214 ± 104.8; n = 3, n = 4, n = 3 biological repeats).

Article Snippet: Materials & experimental systems n/a Involved in the study Antibodies Eukaryotic cell lines Palaeontology and archaeology Animals and other organisms Human research participants Clinical data Dual use research of concern Methods n/a Involved in the study ChIP-seq Flow cytometry MRI-based neuroimaging Antibodies Antibodies used Rabbit-PTBP1 (Abclonal; Cat# - A6107; IF concentration - 1:1000); Mouse-GFAP (Dako; Cat# - M0761; IF concentration - 1:500); Mouse-NeuN (Millipore; Cat# - MAB377; IF concentration - 1:100); Rabbit-pan-ASO (Ionis pharmaceutical, homemade antibody; Cat# -13545; IF concentration - 1:200); Chicken-MAP2 (Abcam; Cat# - ab5392; IF concentration - 1:1000); Mouse-Nestin (abclonal; Cat# - AB22035; IF concentration - 1:200); Rabbit-GFAP (Novus Biologicals; Cat# - NB300-141; IF concentration - 1:1000); Mouse-TubIII (Tuj1) (Millipore Sigma; Cat# - T8660: IF concentration - 1:500); Goat-DCX (Santa Cruz Biotechnology; Cat# - Sc-8066; IF concentration - 1:500).

Techniques: Expressing, Fluorescence, Staining, Labeling, Injection, Two Tailed Test, Control

Journal: PLoS ONE

Article Title: Chronic oral application of a periodontal pathogen results in brain inflammation, neurodegeneration and amyloid beta production in wild type mice

doi: 10.1371/journal.pone.0204941

Figure Lengend Snippet: (A) Confocal microscopy showing e xtracellular Aβ 42 and intracellular Aβ 42 in CA1 region of hippocampus. (B) and (C) are insets from (A). Red arrows in (C) point to intracellular Aβ 42 (Yellow). Green: astrocytes (GFAP+), Red: Aβ 42 detected by rabbit monoclonal antibody, Blue: DAPI. Images are representative of experimental mice (N = 4). (D-F) Images from confocal microscopy of cortex showing intracellular Aβ 42 in 2-D (D), 3-D (E) and orthogonal analysis (F). (G-I) Images from confocal microscopy of hippocampus showing extracellular Aβ 42 in 2-D (G), 3-D (H), and orthogonal analysis (I). Projections a and b are side views of serial confocal sections of the same area.

Article Snippet: Cell and nuclear membrane permeablization was performed by incubating sections in 0.25% Tween 20 in PBS for 30 min. Tissue sections were then independently incubated with 1) mouse monoclonal antibody to Aβ 42 (57001,QED Bioscience, San Diego, CA) and rabbit monoclonal antibody to Aβ 42 (H31L21) (700254, Thermo Fisher Scientific) both at a 1:100 dilution, 2) mouse monoclonal antibody to NeuN (AB 104224, Abcam, Cambridge, MA) and rabbit monoclonal antibody to NeuN (AB177487, Abcam) at a 1:100 dilution to detect intact neurons, 3) goat antibody to Iba1 (NB100, Novus Biologicals, LLC, Littleton, CO) to detect microglia, and 4) mouse monoclonal antibody to anti-GFAP (3670, Cell Signaling Technology, Inc. Danvers, MA) to detect astrocytes at a 1:100 dilution overnight at 4°C.

Techniques: Confocal Microscopy

Journal: Molecular medicine reports

Article Title: Protective effects of tetrahydropalmatine against ketamine-induced learning and memory injury via antioxidative, anti-inflammatory and anti-apoptotic mechanisms in mice.

doi: 10.3892/mmr.2018.8700

Figure Lengend Snippet: Figure 8. Protective effects of THP against GFAP, cytochrome c and PLC‑γ1 protein expression in ketamine‑induced mice. (A) THP protected against GFAP, cytochrome c and PLC‑γ1 protein expression, as determined using western blot analysis. (B‑D) Protein expression levels of GFAP, cytochrome c and PLC‑γ1 in mice treated with ketamine were statistically analyzed. **P<0.05 vs. the control group; ##P<0.05 vs. the model group. GFAP, glial fibrillary acidic protein; PLC‑γ1, phospholipase C‑γ1; THP, tetrahydropalmatine.

Article Snippet: The membranes were blocked with 5% non‐fat milk in TBST for 1 h at 37 ̊C and then incubated with antibodies against iNOS (13120; 1:2,000), GFAP (12389; 1:2,000), GDNF (3897, 1:2,000), cytochrome c (11940; 1:2,000), PLC‐γ1 (5690; 1:2,000) and GAPDH (5174; 1:1,000; all from Cell Signaling Technology, Inc., Danvers, MA, USA) at 4 ̊C overnight.

Techniques: Expressing, Western Blot, Control