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: Stem Cells International

Article Title: Dexamethasone Provides Effective Immunosuppression for Improved Survival of Retinal Organoids after Epiretinal Transplantation

doi: 10.1155/2019/7148032

Figure Lengend Snippet: Primers.

Article Snippet: Antibodies against the following markers were used as primary antibodies for immunofluorescence microscopy at the dilutions indicated in parentheses: Ki67 (1 : 100, A11390, ABclonal, China), CHX10 (1 : 200, AB9016, Millipore, Germany), Brn3 (1 : 200, SC-6026X, Santa Cruz, USA), Islet1 (1 : 20, BM44446, Boster, China), HuD (1 : 100, SC-48421, Santa Cruz), RBPMS (1 : 100, ab152101, Abcam), β III-tubulin (1 : 200, ab7751, Abcam), neurofilament light polypeptide (NEFL; 1 : 100, A0257, ABclonal), microtubule-associated protein 2 (MAP2, 1 : 100, BM1243, Boster), vimentin (1 : 50, BM4029, Boster), glial fibrillary acidic protein (GFAP; 1 : 500, 3670S, Cell Signaling Technology, USA), CD68 (1 : 200, ab201340, Abcam), Iba1 (1 : 200, ab5076, Abcam), and SC121 (1 : 500, Y40410, Takara Bio).

Techniques:

Journal: Stem Cells International

Article Title: Dexamethasone Provides Effective Immunosuppression for Improved Survival of Retinal Organoids after Epiretinal Transplantation

doi: 10.1155/2019/7148032

Figure Lengend Snippet: Characteristics of retinal organoids after 30-45 days of induction. The frozen sections of retinal organoids were identified by immunofluorescence staining. (a, b) At this time, most cells of the organoids were differentiating towards RGCs; they were Islet1-, Brn3- and HuD-positive. (c) But some cells outside were still retinal progenitors; they were CHX10-positive. (d, e) And in suspension cultivation, little axons were developed.

Article Snippet: Antibodies against the following markers were used as primary antibodies for immunofluorescence microscopy at the dilutions indicated in parentheses: Ki67 (1 : 100, A11390, ABclonal, China), CHX10 (1 : 200, AB9016, Millipore, Germany), Brn3 (1 : 200, SC-6026X, Santa Cruz, USA), Islet1 (1 : 20, BM44446, Boster, China), HuD (1 : 100, SC-48421, Santa Cruz), RBPMS (1 : 100, ab152101, Abcam), β III-tubulin (1 : 200, ab7751, Abcam), neurofilament light polypeptide (NEFL; 1 : 100, A0257, ABclonal), microtubule-associated protein 2 (MAP2, 1 : 100, BM1243, Boster), vimentin (1 : 50, BM4029, Boster), glial fibrillary acidic protein (GFAP; 1 : 500, 3670S, Cell Signaling Technology, USA), CD68 (1 : 200, ab201340, Abcam), Iba1 (1 : 200, ab5076, Abcam), and SC121 (1 : 500, Y40410, Takara Bio).

Techniques: Immunofluorescence, Staining, Suspension

Journal: Stem Cells International

Article Title: Dexamethasone Provides Effective Immunosuppression for Improved Survival of Retinal Organoids after Epiretinal Transplantation

doi: 10.1155/2019/7148032

Figure Lengend Snippet: Comparisons of different marker expression profiles among the control, rapamycin- (RAP-) treated, and dexamethasone- (DEX-) treated groups. Two weeks after immunosuppressant treatment, (a) cells in the retinal organoids treated with RAP expressed higher levels of the retinal ganglion cell (RGC) markers Atoh7, Islet1, and Brn3b, the dendrite marker MAP2, the retinal progenitor marker PAX6 and CHX10, and photoreceptor marker CRX, compared to levels in the control group. These cells also expressed lower levels of the axon markers TUBB3 and NEFL than cells in the control group. In DEX-treatment group cells, dendrite, retinal progenitor, and photoreceptor markers (MAP2, CHX10, and CRX) were upregulated compared to levels in the control group. (b) However, 4 weeks after immunosuppressant treatment, both RAP- and DEX-treated cells showed higher expression levels of only PAX6 and CRX as compared to those in the control group. (c) Representative histogram of flow cytometry results. Each wave represented one group; the proportion of each group was calculated by the Blk group. There were 79.87 ± 5.92% of the RGCs in the control group, 60.97 ± 7.36% in the RAP group, and 63.6 ± 10.35% in the DEX group. CTRL: control group; RAP: rapamycin-treatment group; DEX: dexamethasone-treatment group; iPSCs: negative control group in RT-PCR; Blk: blank control group in flow cytometry.

Article Snippet: Antibodies against the following markers were used as primary antibodies for immunofluorescence microscopy at the dilutions indicated in parentheses: Ki67 (1 : 100, A11390, ABclonal, China), CHX10 (1 : 200, AB9016, Millipore, Germany), Brn3 (1 : 200, SC-6026X, Santa Cruz, USA), Islet1 (1 : 20, BM44446, Boster, China), HuD (1 : 100, SC-48421, Santa Cruz), RBPMS (1 : 100, ab152101, Abcam), β III-tubulin (1 : 200, ab7751, Abcam), neurofilament light polypeptide (NEFL; 1 : 100, A0257, ABclonal), microtubule-associated protein 2 (MAP2, 1 : 100, BM1243, Boster), vimentin (1 : 50, BM4029, Boster), glial fibrillary acidic protein (GFAP; 1 : 500, 3670S, Cell Signaling Technology, USA), CD68 (1 : 200, ab201340, Abcam), Iba1 (1 : 200, ab5076, Abcam), and SC121 (1 : 500, Y40410, Takara Bio).

Techniques: Marker, Expressing, Control, Flow Cytometry, Negative Control, Reverse Transcription Polymerase Chain Reaction

Journal: Stem Cells International

Article Title: Isolation of an ES-Derived Cardiovascular Multipotent Cell Population Based on VE-Cadherin Promoter Activity

doi: 10.1155/2016/8305624

Figure Lengend Snippet: Transplantation of CEDPs in adult rats' heart. (a) Expression of Isl1 and MLC2v after 3, 7, and 14 days of transplantation by RT-PCR analysis. mRNA from saline-injected immunosuppressed (N1) or untreated (N2) LV of adult rats was used as negative controls and mRNA from E12 mouse embryos (P) as positive control. (b–d) Clusters of Isl1 + cells derived from CEDPs were detected in frozen sections of LVs isolated 3, 7, and 14 days after transplantation. Sections were stained for DNA with Draq5 and with anti-N-cadherin (b) and anti-cTnT (c). Sections from isolated LVs of saline-injected immunosuppressed rats were used as control for each time-point. (e) All Isl1 + cells from two nonsequential sections were photographed (as in representative upper image) and counted with Fiji cell-counter. Draq5 used for DNA staining. The total Isl1 + cell number found in 2.8 mm tissue was calculated after spline interpolation. Scale bar: 20 μ m.

Article Snippet: For immunocytochemistry, the following antisera were used: rat monoclonals against VE-cadherin (11D4.1, BD Biosciences), PECAM-1 (MEC 13.3, Santa Cruz), and E-Cadherin (DECMA-1, Santa Cruz), mouse monoclonals against cardiac Troponin T (CT3, Iowa Hybridoma Bank), Isl1 (39.4D5, Iowa Hybridoma Bank), Oct3/4 (C-10, Santa Cruz), SMA (Neomarkers), N-cadherin (clone 3B9, Invitrogen), MyHC (MF20, Iowa Hybridoma Bank), and a-actinin (Clone BM-75.2, Sigma), goat polyclonals against GATA4 (C-20, Santa Cruz) and Isl1 (GT15051-100, Acris Antibodies), rabbit monoclonals against MEF2c (D80C1) and VEGF receptor 2 (Flk1) (55B11) from Cell Signaling, and rabbit polyclonals against EGFP (kindly provided from Dr. Charalambia Boleti, Pasteur Institute, Athens), Desmoplakin 1/2 [ ], and DSC2 (DSC2, RDI Research Diagnostics, Inc.).

Techniques: Transplantation Assay, Expressing, Reverse Transcription Polymerase Chain Reaction, Injection, Positive Control, Derivative Assay, Isolation, Staining

Journal: Stem Cells International

Article Title: Isolation of an ES-Derived Cardiovascular Multipotent Cell Population Based on VE-Cadherin Promoter Activity

doi: 10.1155/2016/8305624

Figure Lengend Snippet: Selection of cells with Pvec activity during differentiation. (a) Schematic representation of selection strategy during differentiation of genetically modified ESCs to isolate Pvec + cells. (b) Optical microscope images of clone A11- and E14T-derived cells, during selection at d4 (before the addition of puromycin), d5, and d10 (selected cells). (c) Quantification of EGFP + cells during clone G11 differentiation/selection at d4.5, d6, d7, and d8 by FACS analysis (representative results are shown). (d) Analysis of FACS data from three independent experiments. ∗∗∗ P < 0.001. (e–g) G11-derived EGFP + selected cells at d7 coexpress VE-cadherin, Isl1, and GATA4. Note that EGFP was detected at low Isl1 and GATA4 expressors. (h) Expression of mesodermal, endodermal, and neuroectodermal markers during selection of clone A11 at d3, d7, and d10 by RT-PCR analysis.

Article Snippet: For immunocytochemistry, the following antisera were used: rat monoclonals against VE-cadherin (11D4.1, BD Biosciences), PECAM-1 (MEC 13.3, Santa Cruz), and E-Cadherin (DECMA-1, Santa Cruz), mouse monoclonals against cardiac Troponin T (CT3, Iowa Hybridoma Bank), Isl1 (39.4D5, Iowa Hybridoma Bank), Oct3/4 (C-10, Santa Cruz), SMA (Neomarkers), N-cadherin (clone 3B9, Invitrogen), MyHC (MF20, Iowa Hybridoma Bank), and a-actinin (Clone BM-75.2, Sigma), goat polyclonals against GATA4 (C-20, Santa Cruz) and Isl1 (GT15051-100, Acris Antibodies), rabbit monoclonals against MEF2c (D80C1) and VEGF receptor 2 (Flk1) (55B11) from Cell Signaling, and rabbit polyclonals against EGFP (kindly provided from Dr. Charalambia Boleti, Pasteur Institute, Athens), Desmoplakin 1/2 [ ], and DSC2 (DSC2, RDI Research Diagnostics, Inc.).

Techniques: Selection, Activity Assay, Genetically Modified, Microscopy, Derivative Assay, Expressing, Reverse Transcription Polymerase Chain Reaction

Journal: Stem Cells International

Article Title: Isolation of an ES-Derived Cardiovascular Multipotent Cell Population Based on VE-Cadherin Promoter Activity

doi: 10.1155/2016/8305624

Figure Lengend Snippet: Expansion of Pvec + cells. (a-b) Schematic representation of expansion strategy and propagation of Pvec + cells. Addition of GSK3 inhibitor (SB-216763) induces propagation of Pvec + cells expressing Isl1 at d12. (c) Pvec + /Isl1 + cells percentage in SB-216763 from three independent experiments. For each experiment ~1000 cells were counted. (d) Pvec + cell growth curve. (e–j) Propagated cells consist of cell populations expressing markers of cardiac (Isl1, Mef2c, and GATA4) and/or endothelial (Flk-1, PECAM-1, and VE-cadherin) progenitors.

Article Snippet: For immunocytochemistry, the following antisera were used: rat monoclonals against VE-cadherin (11D4.1, BD Biosciences), PECAM-1 (MEC 13.3, Santa Cruz), and E-Cadherin (DECMA-1, Santa Cruz), mouse monoclonals against cardiac Troponin T (CT3, Iowa Hybridoma Bank), Isl1 (39.4D5, Iowa Hybridoma Bank), Oct3/4 (C-10, Santa Cruz), SMA (Neomarkers), N-cadherin (clone 3B9, Invitrogen), MyHC (MF20, Iowa Hybridoma Bank), and a-actinin (Clone BM-75.2, Sigma), goat polyclonals against GATA4 (C-20, Santa Cruz) and Isl1 (GT15051-100, Acris Antibodies), rabbit monoclonals against MEF2c (D80C1) and VEGF receptor 2 (Flk1) (55B11) from Cell Signaling, and rabbit polyclonals against EGFP (kindly provided from Dr. Charalambia Boleti, Pasteur Institute, Athens), Desmoplakin 1/2 [ ], and DSC2 (DSC2, RDI Research Diagnostics, Inc.).

Techniques: Expressing

Journal: Stem Cells International

Article Title: Isolation of an ES-Derived Cardiovascular Multipotent Cell Population Based on VE-Cadherin Promoter Activity

doi: 10.1155/2016/8305624

Figure Lengend Snippet: Characterization of Pvec + cells. (a) Isl1, GATA4, and VE-cadherin mRNA levels quantification in Pvec + cells in the presence or absence of SB-216763 by real time qPCR. Results in -SB first and -SB second referred to two independent selection/differentiation experiments. (b-c) Isl1 + cardiac progenitors express desmin but not cTnT, a marker of differentiated cardiomyocytes. (d) RT-PCR analysis of expanded Pvec + cells showed expression of cardiac, endothelial, and endocardial but not neuroectodermal, endodermal, or pluripotency markers. As positive controls (ctr) mRNA from E14T ESCs differentiation was used. Note the lack of cTnT, MLC2v, and MLC2a expression in expanded Pvec + cells. Scale bar: 20 μ m.

Article Snippet: For immunocytochemistry, the following antisera were used: rat monoclonals against VE-cadherin (11D4.1, BD Biosciences), PECAM-1 (MEC 13.3, Santa Cruz), and E-Cadherin (DECMA-1, Santa Cruz), mouse monoclonals against cardiac Troponin T (CT3, Iowa Hybridoma Bank), Isl1 (39.4D5, Iowa Hybridoma Bank), Oct3/4 (C-10, Santa Cruz), SMA (Neomarkers), N-cadherin (clone 3B9, Invitrogen), MyHC (MF20, Iowa Hybridoma Bank), and a-actinin (Clone BM-75.2, Sigma), goat polyclonals against GATA4 (C-20, Santa Cruz) and Isl1 (GT15051-100, Acris Antibodies), rabbit monoclonals against MEF2c (D80C1) and VEGF receptor 2 (Flk1) (55B11) from Cell Signaling, and rabbit polyclonals against EGFP (kindly provided from Dr. Charalambia Boleti, Pasteur Institute, Athens), Desmoplakin 1/2 [ ], and DSC2 (DSC2, RDI Research Diagnostics, Inc.).

Techniques: Selection, Marker, Reverse Transcription Polymerase Chain Reaction, Expressing

Journal: Stem Cells International

Article Title: Isolation of an ES-Derived Cardiovascular Multipotent Cell Population Based on VE-Cadherin Promoter Activity

doi: 10.1155/2016/8305624

Figure Lengend Snippet: CEDPs differentiation potential. (a) Optical microscope image of a sphere formed in the absence of SB-216763 after 5 days of CEDPs differentiation. (b) cTnT + cells formation after immunostaining with anti-cTnT and anti-VE-cadherin. (c–e) Formation of intercalated disk structures in beating spheres after 10 days of CEDPs differentiation, shown by double-IF-staining with anti-cTnT/anti-Desmoplakin (DSP), anti-cTnT/anti-Desmocollin-2 (DSG2), and anti-MyHC/anti-Desmoplakin. Note the striations of the sarcomeric cTnT staining and the punctate desmosomal staining in areas connecting adjacent cardiomyocytes. (f) Expression of MLC2v and MLC2a after 10 days of CEDPs differentiation by RT-PCR analysis. (g) VE-cadherin + cells in CEDPs-derived differentiated cells. Note extensive adherens junctions formation between endothelial cells. Magnification corresponds to marked area. (h-i) Expression of vWF and CD39 in VE-cadherin + endothelial cells after 10 days of CEDPs differentiation. (j-k) Expression of Isl1 and Mef2c progenitor markers in a-actinin + and VE-cadherin + cells, respectively, at day 10 of CEDPs differentiation. (l) Expression of SMA during CEDPs differentiation. Draq5 counterstained DNA (b, g). Scale bar: 20 μ m.

Article Snippet: For immunocytochemistry, the following antisera were used: rat monoclonals against VE-cadherin (11D4.1, BD Biosciences), PECAM-1 (MEC 13.3, Santa Cruz), and E-Cadherin (DECMA-1, Santa Cruz), mouse monoclonals against cardiac Troponin T (CT3, Iowa Hybridoma Bank), Isl1 (39.4D5, Iowa Hybridoma Bank), Oct3/4 (C-10, Santa Cruz), SMA (Neomarkers), N-cadherin (clone 3B9, Invitrogen), MyHC (MF20, Iowa Hybridoma Bank), and a-actinin (Clone BM-75.2, Sigma), goat polyclonals against GATA4 (C-20, Santa Cruz) and Isl1 (GT15051-100, Acris Antibodies), rabbit monoclonals against MEF2c (D80C1) and VEGF receptor 2 (Flk1) (55B11) from Cell Signaling, and rabbit polyclonals against EGFP (kindly provided from Dr. Charalambia Boleti, Pasteur Institute, Athens), Desmoplakin 1/2 [ ], and DSC2 (DSC2, RDI Research Diagnostics, Inc.).

Techniques: Microscopy, Immunostaining, Staining, Expressing, Reverse Transcription Polymerase Chain Reaction, Derivative Assay

Journal: Stem Cells International

Article Title: Isolation of an ES-Derived Cardiovascular Multipotent Cell Population Based on VE-Cadherin Promoter Activity

doi: 10.1155/2016/8305624

Figure Lengend Snippet: Characterization of CEDPs-derived products. (a) Statistical analysis of cTnT + and VE-cadherin + cells in CEDPs-derived differentiated cells at d12. The total number of cells counted was 10417 in three independent experiments. ∗∗∗ P < 0.001 and ∗∗ P < 0.01. Cells from random fields were photographed (as in representative image left) and counted by Fiji cell-counter. Scale bar: 20 μ m. (b) cTnT, Nkx2.5, Isl1, GATA4, VE-cadherin, and Flk1 mRNA levels quantification in CEDPs-derived differentiated cells at d10 by real time qPCR. Results in CEDPs differentiation first and CEDPs differentiation second referred to two independent differentiation experiments.

Article Snippet: For immunocytochemistry, the following antisera were used: rat monoclonals against VE-cadherin (11D4.1, BD Biosciences), PECAM-1 (MEC 13.3, Santa Cruz), and E-Cadherin (DECMA-1, Santa Cruz), mouse monoclonals against cardiac Troponin T (CT3, Iowa Hybridoma Bank), Isl1 (39.4D5, Iowa Hybridoma Bank), Oct3/4 (C-10, Santa Cruz), SMA (Neomarkers), N-cadherin (clone 3B9, Invitrogen), MyHC (MF20, Iowa Hybridoma Bank), and a-actinin (Clone BM-75.2, Sigma), goat polyclonals against GATA4 (C-20, Santa Cruz) and Isl1 (GT15051-100, Acris Antibodies), rabbit monoclonals against MEF2c (D80C1) and VEGF receptor 2 (Flk1) (55B11) from Cell Signaling, and rabbit polyclonals against EGFP (kindly provided from Dr. Charalambia Boleti, Pasteur Institute, Athens), Desmoplakin 1/2 [ ], and DSC2 (DSC2, RDI Research Diagnostics, Inc.).

Techniques: Derivative Assay

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