Journal: Molecular Genetics and Metabolism Reports

Article Title: Generation of GLA-knockout human embryonic stem cell lines to model peripheral neuropathy in Fabry disease

doi: 10.1016/j.ymgmr.2022.100914

Figure Lengend Snippet: Antibodies used for immunostaining.

Article Snippet: Islet1/ISL1 , Islet1/ISL1 , rabbit IgG , 1:500 , Sino Biological , Wayne, PA.

Techniques: Immunostaining

Journal: Molecular Genetics and Metabolism Reports

Article Title: Generation of GLA-knockout human embryonic stem cell lines to model peripheral neuropathy in Fabry disease

doi: 10.1016/j.ymgmr.2022.100914

Figure Lengend Snippet: Immunostaining of neuronal cells derived from AGA-deficient WA14 clones for markers of sensory neurons. WA14 clones were differentiated as described in Methods. At Day 8, they were transferred to Matrigel-coated 2-well glass chamber slides and the differentiation protocol was continued to Day 12, after which the cells were refed with N2 growth medium. At Day 14, the cultures were fixed and stained as described in Methods. (A) Cells were double stained for peripherin (green) and beta-III-tubulin (TUBB3, red). Areas of overlap appear as yellow. (B) Cells were double stained for BRN3A (green) and beta-III-tubulin (TUJ1, red). (C) Cells were double stained for Islet1 (green) and beta-III-tubulin (TUJ1, red). All slides were imaged in a Keyence 9000 microscope using a 20× objective. Scale bars = 100 μm.

Article Snippet: Islet1/ISL1 , Islet1/ISL1 , rabbit IgG , 1:500 , Sino Biological , Wayne, PA.

Techniques: Immunostaining, Derivative Assay, Clone Assay, Staining, Microscopy

Journal: bioRxiv

Article Title: Decreased CREB phosphorylation impairs embryonic retinal neurogenesis in the Oa1-/- mouse model of Ocular albinism

doi: 10.1101/2024.05.14.594013

Figure Lengend Snippet: (A) Images show Oa1-/- retina has larger nuclei (asterisks), fewer Tuj1+ cells and reduced Tuj1 intensity per unit area, respectively. The boxed areas show zoomed retinal cells. Bars, 50 μm. (B) qRT-PCR showing reduced Tfap2b and Dcx mRNAs in E15.5 Oa1-/- eyes compared to WT. (C) Reduced Tfap2b (AC marker) expression in E15.5 Oa1-/- mouse eyes and retinas (yellow arrowheads). Bars, 50 μm. (D) Western blot and bar graphs show reduced Tfap2b level in Oa1-/- eyes. pCREB and pATF1 levels are also reduced in Oa1-/- eyes. n=6-12 E15.5 littermates, 3 litters/group. (E) Representative image shows Dcx+ RGCs (red) in the central WT retina. (F) Dcx+ RGCs from E15.5 retinas are non-proliferating (Ki67-) and express Islet1, a marker of differentiated RGCs. Ki67 staining is in a different section than that shown in . (G) E15.5 eyes show an expansion in Dcx (green) staining. The zoomed images (orientation changed) show reduction of Dcx in Oa1-/- mouse retinas (yellow arrowheads). The length of the Dcx labeled neurites (black line bar) is shorter in Oa1-/- retinas. Co-labelling for Dcx (red) and pCREB (green) shows both markers decreased in Oa1-/- eyes. DAPI, nuclei. (H) Western blot and bar graphs show reduced Dcx level in E15.5 Oa1-/- eyes. n=6-12 littermates, 3 litters/group. Significance was determined by Holms-Sidak tests, except in 4A and 4B, analyzed by Welch’s t- tests. (I) Schematic representation of Oa1-induced CREB activation in an embryonic mouse eye. RPE-specific Oa1 signals mainly through Epac1-pErk2 to phosphorylate CREB in the RPE. This is followed by CREB activation in the adjacent retina, leading to proliferation and differentiation.

Article Snippet: List of primary antibodies used: Rabbit anti-GPR143 (1:500, Sigma, Cat#HPA003648), Rabbit anti-Gα s (1:100; Abcam, Cat#ab83735), Rabbit anti-Epac1(1:100, Abcam, Cat#ab21236), Rabbit anti-phospho CREB (1:100, Cell Signaling, Cat#9198), Rabbit anti-phospho Erk1/2 (1:100, Cell Signaling, Cat#9101), Rabbit anti-Dcx (1:200, Abcam, Cat#ab6142), Rabbit anti-AP-2beta (1:1200, Cell Signaling, Cat#2509), Rabbit anti-Ki67 (1:1000, Abcam, Cat#ab15580), Rabbit anti-Islet1 (1: 500, Abcam, Cat#ab20670), Mouse anti-Nestin (1:100, Abcam, Cat#ab6142), Mouse anti-Tuj1 (1:500, Abcam, Cat#ab78078), Mouse anti-CyclinD2 (1:50, SCBT, Cat#sc-56305).

Techniques: Quantitative RT-PCR, Marker, Expressing, Western Blot, Staining, Labeling, Activation Assay

Journal: Cell Reports Medicine

Article Title: Targeting low levels of MIF expression as a potential therapeutic strategy for ALS

doi: 10.1016/j.xcrm.2024.101546

Figure Lengend Snippet:

Article Snippet: Primary antibodies (mouse monoclonal anti-beta III tubulin (Tuj1) 1:1000; rabbit monoclonal Tuj1 1:1000; rabbit polyclonal anti-Map 2 1:1000; rabbit polyclonal anti-Islet1 1:1000; rabbit monoclonal anti-MIF 1:500 all from Abcam, Cambridge, UK and mouse monoclonal anti-Misfolded SOD1 (B8H10), Biomol GmbH (Medimabs), Hamburg, Germany) were diluted in blocking solution and incubated overnight at 4°C.

Techniques: Virus, Recombinant, SYBR Green Assay, Bicinchoninic Acid Protein Assay, Software, Real-time Polymerase Chain Reaction, RNA Sequencing Assay

Journal: iScience

Article Title: Preclinical long-term safety of intraspinal transplantation of human dorsal spinal GABA neural progenitor cells

doi: 10.1016/j.isci.2023.108306

Figure Lengend Snippet:

Article Snippet: Rabbit monoclonal anti-ISLET1 , Abcam , Cat#ab109517; RRID: AB_10866454.

Techniques: Recombinant, Blocking Assay, Software, Microscopy, Laser-Scanning Microscopy