Journal: Neuron

Article Title: Sox11 Expression Promotes Regeneration of Some Retinal Ganglion Cell Types but Kills Others

doi: 10.1016/j.neuron.2017.05.035

Figure Lengend Snippet: (A, B) Representative images of optic nerve sections showing CTB-labeled axons in wild type mice with intravitreal injections of AAV expressing PLAP, Sox4, and Sox11 (A), and Pax6, Math5, Sox2, Brn3b, and Isl1 (B) at 2 weeks after optic nerve injury. The crush site is indicated with a red asterisk. Scale bars in (A) and (B) represent 250 µm.

Article Snippet: In vivo procedures and reagents Production of AAVs Vectors of AAV-Pax6 and AAV-Isl1 vectors were made by inserting mouse Pax6 cDNA (Addgene #32932) and mouse Isl1 cDNA (Addgene #32929) respectively into an AAV plasmid consisting of the cytomegalovirus enhancer fused to the chicken beta-actin promoter (CAG promoter) and Woodchuck hepatitis virus Posttranscriptional Regulatory Element (WPRE).

Techniques: Labeling, Expressing

Journal: International Journal of Ophthalmology

Article Title: Nintedanib induces apoptosis in human pterygium cells through the FGFR2-ERK signalling pathway

doi: 10.18240/ijo.2023.04.03

Figure Lengend Snippet: A: Phase contrast microscopy of primary human pterygium-derived cells; B: Phase contrast microscopy of passaged human pterygium-derived cells (Generation 8); C: The results showed that the purity of pterygium epithelial cells accounted for up to 74%. MUC1: transmembrane glycoprotein mucin 1; K10: Keratin 10.

Article Snippet: The cells were then incubated in the dark for 30min at 4°C with FITC-labeled MUC1 (559774, BD) and PE-labeled K10 (orb485164, Biorbyt) antibodies.

Techniques: Microscopy, Derivative Assay

Journal: Frontiers in bioscience (Landmark edition)

Article Title: Single-Cell Transcriptome Analysis of Small Cell Neuroendocrine Carcinoma of the Endometrium Reveals ISL1 as a Potential Biomarker for Diagnosis and Treatment.

doi: 10.31083/j.fbl2903100

Figure Lengend Snippet: Fig. 5. ISL1 modulates neuroactive pathway in subclusters of single cell RNA-seq data and The Cancer Genome Atlas (TCGA)

Article Snippet: After cell counting, the cells were incubated with ISL LIM Homeobox 1 (ISL1) monoclonal antibody (1:200, 15661-1-AP; Proteintech, Rosemont, IL, USA) at 4 °C in the dark for 60 min, followed by incubation for 30 min with a fluorescent secondary antibody (1:200, SA00013-2, CoraLite488-conjugated goat anti-rabbit IgG (H+L)).

Techniques: RNA Sequencing

Journal: Frontiers in bioscience (Landmark edition)

Article Title: Single-Cell Transcriptome Analysis of Small Cell Neuroendocrine Carcinoma of the Endometrium Reveals ISL1 as a Potential Biomarker for Diagnosis and Treatment.

doi: 10.31083/j.fbl2903100

Figure Lengend Snippet: Fig. 6. Flow cytometric sorting of neuroendocrine tumor cells to obtain ISL1 expression group, and explore the differences in cell function in vitro. (A) Flow cytometric sorting of neuroendocrine tumor cell line H446 according to the difference in the expression of

Article Snippet: After cell counting, the cells were incubated with ISL LIM Homeobox 1 (ISL1) monoclonal antibody (1:200, 15661-1-AP; Proteintech, Rosemont, IL, USA) at 4 °C in the dark for 60 min, followed by incubation for 30 min with a fluorescent secondary antibody (1:200, SA00013-2, CoraLite488-conjugated goat anti-rabbit IgG (H+L)).

Techniques: Expressing, Cell Function Assay, In Vitro

Journal: Developmental cell

Article Title: Isl1 regulation of Nkx2.1 in the early foregut epithelium is required for tracheaesophageal separation and lung lobation

doi: 10.1016/j.devcel.2019.11.002

Figure Lengend Snippet: Key Resources Table

Article Snippet: Plasmid: pCMV-Isl1 , OriGene , Cat#MR227038.

Techniques: Recombinant, Reporter Assay, Mutagenesis, Isolation, Microarray, Plasmid Preparation, Software

Journal: Nature communications

Article Title: High-efficiency motor neuron differentiation from human pluripotent stem cells and the function of Islet-1.

doi: 10.1038/ncomms4449

Figure Lengend Snippet: Figure 5 | ISL1 is a critical regulator of human motoneuron development. (a) Immunofluorescence of ISL1 (red) in cells after 15-day differentiation from H1 hESCs. Cell nuclei were stained with DAPI (4’,6-diamidino-2-phenylindole; blue). RA-D1, RA-D3 and RA-D6 denote the time of initiation of RA patterning (at day 1, day 3 and day 6, respectively). (b) Percentage of ISL1-positive cells under conditions described in a. Each bar represents mean±s.d. (error bars) of six experiments. (c) Fluorescence images of ISL1 (left), ISL1/DAPI (middle) and ISL1/ TUJ1 (right) staining in differentiated cells replated at a low density for 3 days after 15-day differentiation. (d) Fluorescence images of differentiated hESCs with NT shRNA (‘Control’, left) or with ISL1 shRNA-1 (‘ISL shRNA-1’, right) stained with antibodies against ISL1 (red) and TUJ1 (green) after 15-day differentiation (with patterning initiated at day 3). Cell nuclei were stained with DAPI (blue). (e) Fluorescence images of differentiated hESCs with NTshRNA (‘Control’, left), ISL1 shRNA-2 (‘ISL shRNA-2’, middle) and ISL1 shRNA-2 with ectopic ISL1 expression (‘shRNA-2 þ ISL1’, right) stained with antibodies against ISL1 (red) and TUJ1 (green) after 15-day differentiation (with patterning initiated at day 3). Cell nuclei were stained with DAPI (blue). (f) Fluorescence images of differentiated hESCs with NTshRNA (left) or with ISL1 shRNA (right) stained with antibodies against HB9 (red) and ChAT (green) after 20-day differentiation. Cell nuclei were stained with DAPI (blue). (g) Fluorescence images of differentiated hESCs with NT shRNA (left), ISL1 shRNA-2 (middle) and ISL1 shRNA-2, with ectopic ISL1 expression (right) stained with antibodies against HB9 (red) and ChAT (green) after 20-day differentiation (with patterning initiated at day 3). Cell nuclei were stained with DAPI (blue). All scale bars, 50 mm.

Article Snippet: For ISL1 rescue experiments, ISL1 expression vector was generated by inserting human ISL1 cDNA (OriGene SC125257) into pSin-EF2-alpha-Puro lentivector (Addgene).

Techniques: Staining, Fluorescence, shRNA, Control, Expressing

Journal: Nature communications

Article Title: High-efficiency motor neuron differentiation from human pluripotent stem cells and the function of Islet-1.

doi: 10.1038/ncomms4449

Figure Lengend Snippet: Figure 6 | Summary of the MN differentiation model and the stages of neural differentiation. (a) hESCs undergoing neural induction begin to express high levels of NPC markers, including PAX6 and SOX1 at day 3, while markedly downregulating the pluripotency markers such as OCT-4 and NANOG (albeit retaining some degree of expression). Thus, the cells at day 3 might represent a population of primitive NPCs that can be induced to become anterior NPCs and also possesses the potential to differentiate posteriorly. With neural patterning initiated at day 3 after neural induction, the early MN marker ISL1 can be seen at day 13 after hESC differentiation, while mature MN markers including HB9 and ChAT are expressed after 17-day differentiation. (b) Our results point to a previously unidentified primitive stage of neural progenitors that is intermediate between pluripotent hESCs and anterior NPCs. This population of potential primitive NPCs can be induced to become anterior NPCs in the absence of patterning factors, and also possesses the potential to differentiate along the posterior fate in the presence of patterning factors.

Article Snippet: For ISL1 rescue experiments, ISL1 expression vector was generated by inserting human ISL1 cDNA (OriGene SC125257) into pSin-EF2-alpha-Puro lentivector (Addgene).

Techniques: Expressing, Marker

Journal: Developmental cell

Article Title: Isl1 regulation of Nkx2.1 in the early foregut epithelium is required for tracheaesophageal separation and lung lobation

doi: 10.1016/j.devcel.2019.11.002

Figure Lengend Snippet: Key Resources Table

Article Snippet: The pCMV-Isl1 plasmid was purchased from OriGene.

Techniques: Recombinant, Reporter Assay, Mutagenesis, Isolation, Microarray, Plasmid Preparation, Software