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: Journal of Clinical Medicine

Article Title: Diagnostic and Prognostic Value of Serum Glial Fibrillary Acidic Protein in Acute Ischemic Stroke

doi: 10.3390/jcm15051971

Figure Lengend Snippet: Serum GFAP levels in patients and controls.

Article Snippet: Serum GFAP levels were measured by the Lumipulse G GFAP assay on the fully automated platform Lumipulse G1200 (FUJIREBIO Inc., Tokyo, Japan).

Techniques:

Journal: Journal of Clinical Medicine

Article Title: Diagnostic and Prognostic Value of Serum Glial Fibrillary Acidic Protein in Acute Ischemic Stroke

doi: 10.3390/jcm15051971

Figure Lengend Snippet: ROC curve analysis of serum GFAP for detecting acute ischemic stroke.

Article Snippet: Serum GFAP levels were measured by the Lumipulse G GFAP assay on the fully automated platform Lumipulse G1200 (FUJIREBIO Inc., Tokyo, Japan).

Techniques:

Journal: Journal of Clinical Medicine

Article Title: Diagnostic and Prognostic Value of Serum Glial Fibrillary Acidic Protein in Acute Ischemic Stroke

doi: 10.3390/jcm15051971

Figure Lengend Snippet: GFAP levels in patients according to TOAST classification.

Article Snippet: Serum GFAP levels were measured by the Lumipulse G GFAP assay on the fully automated platform Lumipulse G1200 (FUJIREBIO Inc., Tokyo, Japan).

Techniques:

Journal: Journal of Clinical Medicine

Article Title: Diagnostic and Prognostic Value of Serum Glial Fibrillary Acidic Protein in Acute Ischemic Stroke

doi: 10.3390/jcm15051971

Figure Lengend Snippet: Correlation analysis between GFAP and NIHSS at different time points. T0, at admission; T6, after six days of hospitalization; and TD, at discharge.

Article Snippet: Serum GFAP levels were measured by the Lumipulse G GFAP assay on the fully automated platform Lumipulse G1200 (FUJIREBIO Inc., Tokyo, Japan).

Techniques:

Journal: Journal of Clinical Medicine

Article Title: Diagnostic and Prognostic Value of Serum Glial Fibrillary Acidic Protein in Acute Ischemic Stroke

doi: 10.3390/jcm15051971

Figure Lengend Snippet: Correlation analysis between GFAP and mRS at different time points. T0, at admission; T6, after six days of hospitalization.

Article Snippet: Serum GFAP levels were measured by the Lumipulse G GFAP assay on the fully automated platform Lumipulse G1200 (FUJIREBIO Inc., Tokyo, Japan).

Techniques:

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Tissue- and cell-type-specific manifestations of heteroplasmic mtDNA 3243A>G mutation in human induced pluripotent stem cell-derived disease model

doi: 10.1073/pnas.1311660110

Figure Lengend Snippet: Characterization of MELAS iPSCs. (A) m.3243A>G MELAS mutant mtDNA amount in parental fibroblast cultures (Fb), the reprogrammed iPSC lines (iPSC), and in fibroblast clones from the parental lines (Fb_cl). (B) RT-PCR analysis of ES cell-specific transcripts (OCT4, SOX2, NANOG, REX, and DNMT3B). All patient-derived iPSC clones expressed ES-specific genes similarly to the healthy control iPSCs and ES cells (HDF, human dermal fibroblasts; HES, human embryonic stem cells). (C) Immunofluorescence staining for ES cell-marker proteins TRA-1–60 (green), SSEA4 (green), and NANOG (red). Blue staining for nucleus is DAPI. Colony morphology and ES cell-marker protein expression were similar in all of the clones. (Scale bar, 50 μm.) (D) Expression of OCT4, SOX2, KLF4, and c-MYC compared with that in human ES cells and normalized against cyclophilin, expression. Expression of viral transgenes was down-regulated in all clones. (E) MELAS iPSC lines generated teratomas that differentiated toward all three germ layers independent of the mutation load. (Left) Mesoderm, cartilage; (Center) ectoderm, pigmented epithelia; and (Right) endoderm, intestinal epithelia. (Scale bar, 200 μm.) (F) iPSCs differentiated in vitro into neural cultures consisting of MAP2 and βIII-tubulin positive neurons (green) and GFAP-positive glia (red). DAPI staining for nuclei (blue). (Scale bar, 50 μm.) (G) m.3243A>G mutant mtDNA load in the iPSC lines during culture and in the differentiated cells. The heteroplasmy levels did not change significantly during culture or upon differentiation. MH, MELAS-high; ML, MELAS-low; p., passage number. See also Fig. S1.

Article Snippet: To reveal antigenic sites, the rehydrated sections were treated in a microwave in 1 mM EDTA, pH 8, for 3 min. Primary antibodies were against Tra1-60 (1:40, ab90232, Millipore), SSEA4 (1:1,000, ab90231, Millipore), Nanog (1:500, D73G4, Cell Signaling Technology), β-III-tubulin [1:100 (immunofluorescence, IF), 1:500 (immunohistochemistry, IHC), T2200, Sigma], MAP2 (1:2,000, ab5543, Millipore), GFAP (1:100, mab2594, R&D Systems), DNA (1:20, AC-30-10, Progen Biotechnik), GM130 (1:250, sc16268, Santa Cruz Biotechnology), COX1 [1:200 (IF), 1:500 (western blotting, WB), M404, Mitosciences], C1-NDUFS3 (1:142, ab110246, abcam), CI-NDUFB4 (1:2,000, ab110243, abcam), CI-NDUFA9 (1:2,000, ab14713, abcam), CII-70kDa [1:2,000 (IF), 1:10000 (WB), 1:142 (IHC), MS204, Mitosciences], CIII-UQCRC2 [1:2,500 (WB), 1:1,000 (IF), 1:142 (IHC), ab14745, abcam], CV-α (1:1,000, MS507, Mitosciences), LC3B (1:200, NB600-1384, Novus Biologicals), PINK1 [1:250 (IF), 1:500 (WB), ab75487, abcam], PRK8 1:250 (IF), 1:1,000 [(WB) ab15954, abcam], β-tubulin (1:5,000, 2146, Cell Signaling Technology), β-Actin (1:2,000, sc1616, Santa Cruz Biotechnology), and TOM20 [1:250 (IF), 1:500 (WB), sc11415, Santa Cruz Biotechnology].

Techniques: Mutagenesis, Clone Assay, Reverse Transcription Polymerase Chain Reaction, Derivative Assay, Control, Immunofluorescence, Staining, Marker, Expressing, Generated, In Vitro

Journal: Cell Reports Methods

Article Title: Reliable multiplex generation of pooled induced pluripotent stem cells

doi: 10.1016/j.crmeth.2023.100570

Figure Lengend Snippet:

Article Snippet: Mouse monoclonal anti-human GFAP , Novus Biologicals , Cat#NBP2-33184AF647; RRID: AB_2935766.

Techniques: Recombinant, Membrane, Flow Cytometry, Electron Microscopy, DNA Extraction, Sample Prep, Sequencing, Plasmid Preparation, Software