anti-tra-1-60 Search Results


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  • 95
    Abcam anti tra 1 60
    hAEC are positive for naïve pluripotent markers. (A) Representative micrographs at 20x from hAEC at P0-P2 immunostained for KLF4 (red) and TFE3 (red); the nuclei were stained with DAPI (blue). (B) Graph shows the percentage of hAEC that express naïve markers. (C) Representative micrographs at 20x from hAEC at P0-P2 immunostained for KLF4 (red) or TFE3 (red) and co-expressing <t>TRA-1-60;</t> the nuclei were stained with DAPI (blue). (D) Graph shows the percentage of cells immunostained for both TRA-1-60 and naïve markers. Results are expressed as percentages of means ± S.E.M. from 9 fields counted in duplicate from three independent experiments. Scale bar 50 μm. * p
    Anti Tra 1 60, supplied by Abcam, used in various techniques. Bioz Stars score: 95/100, based on 51 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Millipore anti tra 1 60
    Hsa-let-7c overexpression alters human induced neuronal (iN) cell morphology. (A) induced pluripotent stem (iPS) cells stain positive for pluripotent markers OCT4 and <t>Tra-1-60.</t> (B) iN cell induction protocol for converting human iPS cells into functional glutamatergic neurons. (C) Immunocytochemical characterization of excitatory iN cells. iN cells transduced with Ngn2 stain positive for the neuronal lineage marker TUJ1, mature neuronal marker MAP2, and presynaptic marker synapsin; Ngn2-iN cells are also positive for the excitatory synaptic marker, vGlut1, and negative for the inhibitory marker vGAT. (D) Design for the let-7c overexpression construct. The H1 promoter and a downstream short hairpin RNA (shRNA) containing the mature let-7c sequence were cloned into the FG12 lentiviral backbone. (E) HEK293T cells were transfected with either FG12 or FG12-let-7c and assayed via qPCR to assess let-7c overexpression. (F–H) The FG12-let-7c construct was packaged into lentivirus and used to infect human iN cells according to (B) . Infected cells express GFP (F) and AG2U cells show a significant reduction in soma size (G) , as well as primary dendrite outgrowth (H) . Statistical tests were performed using two-tailed Student’s t -test, where * p
    Anti Tra 1 60, supplied by Millipore, used in various techniques. Bioz Stars score: 93/100, based on 438 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Becton Dickinson anti tra 1 60
    hPSCs cultured in the defined, xeno-free EOD hPSC medium retain their stem cell characteristics. Immunodetection of pluripotency antigens in hPSC lines cultivated for 40 passages in the defined, xeno-free L7 hPSC medium with EOD medium changes. Detection of Oct-3/4 and Nanog are shown (red) along with SSEA-4 and <t>TRA-1-60</t> (green). Individual cell nuclei were visualized using DAPI (blue). Scale bar: 200 μm.
    Anti Tra 1 60, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 91/100, based on 100 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Stemgent anti tra 1 60
    In vitro characterization of the pluripotent state of EiPS cells. ( a ). Nanog, SSEA1, SSEA4, <t>TRA-1-60</t> and TRA-1-81 detected by fluorescent immunohistochemistry. Yellow scale bar 10 μm, white scalebar 20 μm. ( b ). Detection of gene expression of key pluripotency markers using RT-PCR primers specific for equine endogenous genes
    Anti Tra 1 60, supplied by Stemgent, used in various techniques. Bioz Stars score: 91/100, based on 21 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    BioLegend anti tra 1 60
    XPC increases somatic reprogramming fidelity and iPSC self-renewal capacity. ( A . ( B ) Flow cytometry analysis of control and XPC complex (XPC–RAD23B–CETN2) gain-of-function cell populations 24 d post-induction for the late stage human iPSC marker <t>TRA-1-60.</t> ( C ) The average number of colonies obtained in the reprogramming experiment depicted in B . ( D ) iPSCs derived from control or XPC–RAD23B–CETN2-overexpressing HDFs were challenged with single-cell dissociation and allowed to recover for 3–4 d before staining with alkaline phosphatase (AP). The average number of AP + colonies formed per 2.5 × 10 4 single cells plated. Error bars depict the standard deviation. n = 3. (*) P
    Anti Tra 1 60, supplied by BioLegend, used in various techniques. Bioz Stars score: 94/100, based on 13 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Cell Signaling Technology Inc anti tra 1 60
    Characterization of the two types of piPSCs. (A)The colonies of mpiPSCs were round and three-dimensional (bottom). The colonies of hpiPSCs were large and flat (top). Bar scale = 500 μm. The hpiPSCs and mpiPSCs were both positive for alkaline phosphatase (AP). Bar scale = 200 μm. (B) Immunocytochemical staining of Oct4, Sox2, SSEA-1 and SSEA-4 in hpiPSCs and mpiPSCs. Bar scale = 200 μm. (C) The surface marker Tra-1-81 and <t>Tra-1-60</t> were positive in hpiPSCs but not detected in mpiPSCs. Bar scale = 200 μm. (D) X chromosome activation state of hpiPSCs and mpiPSCs after immunostaining for H3K27me3. Positive signals of histone H3K27 me3 spots were observed in hpiPSCs but not in mpiPSCs. Bar scale = 200 μm. The white arrowheads indicate the H3K27me3 positive spot in the cell. (E) Immunocytochemical assay of 3-germ-layer cells in EBs derived from both types of piPSCs, the markers include β Ⅲ-Tubulin (ectoderm), α-SMA (mesoderm) and Sox17 (endoderm). Scale bar = 200 μm. (F) Hematoxylin and eosin staining of teratoma sections of piPSCs. Left: blood vessel of endothelium (ectoderm); middle: muscle (mesoderm); right: gut-like epithelium (endoderm). Scale bars = 200 μm. (G) Karyotype analysis of piPSCs.
    Anti Tra 1 60, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 91/100, based on 78 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Merck KGaA anti tra 1 60
    Confirmation of pluripotent markers in cells and tissue samples. ( a ) Relative gene expression of OCT4. ( b ) Relative gene expression of LIN28 in cells and pellets. ( c ) Fluorescence image of iPSC-derived teratoma tissues stained with the <t>TRA-1-60</t> and c-Myc antibody. Fluorescence image of ( d ) mcMock, ( e ) mcBMP2, ( f ) mcTGFβ3, and ( g ) mcBOTH. Images show the 200× magnification image. Scale bar represents 200 μm. mcMock: minicircle mock vector-transfected outgrowth cells; mcBMP2: minicircle BMP2-encoding vector-transfected outgrowth cells; mcTGFβ3: minicircle TGFβ3-encoding vector-transfected outgrowth cells; mcBOTH: 1:1 mixture of mcBMP2- and mc TGFβ3-transfected outgrowth cells.
    Anti Tra 1 60, supplied by Merck KGaA, used in various techniques. Bioz Stars score: 91/100, based on 17 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Santa Cruz Biotechnology anti tra 1 60
    Undifferentiated ORMES6 ESCs express embryonic cell markers. (A) SSEA 4 (green); (B) Oct4 (red); (C) glycoprotein <t>TRA-1-60</t> (green)/ Oct4 (red) and (D) glycoprotein TRA-1-81 (green)/Oct4 (red). The nuclei were stained with DAPI (blue). The undifferentiated ORMES6 ESCs expressed SSEA 4, Oct4, TRA-1-60 and TRA-1-81 antigens. Cells only stained with DAPI were MEF feeder cells. Scale bar: 40 μm.
    Anti Tra 1 60, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 91/100, based on 36 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Millipore mouse anti tra 1 60
    Verification of the pluripotency of the iPSC lines with the TDP-43 A90V mutation. ( A ) Fluorescence microscopy images of the expression of the pluripotency markers NANOG, OCT4, SSEA4, <t>TRA-1-60,</t> and TRA-1-81 in control (37L25) and patient (36L10) iPSC lines. Scale bar: 20 µm. ( B ) All iPSC lines differentiated into cells of the three germ layers, as shown by expression of desmin (mesoderm), TUJ1 (ectoderm), and alpha-fetoprotein (AFP, endoderm). These analyses indicate iPSC lines generated here are indeed pluripotent. Scale bar: 20 µm.
    Mouse Anti Tra 1 60, supplied by Millipore, used in various techniques. Bioz Stars score: 90/100, based on 243 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Becton Dickinson tra 1 60 alexa 488
    Verification of the pluripotency of the iPSC lines with the TDP-43 A90V mutation. ( A ) Fluorescence microscopy images of the expression of the pluripotency markers NANOG, OCT4, SSEA4, <t>TRA-1-60,</t> and TRA-1-81 in control (37L25) and patient (36L10) iPSC lines. Scale bar: 20 µm. ( B ) All iPSC lines differentiated into cells of the three germ layers, as shown by expression of desmin (mesoderm), TUJ1 (ectoderm), and alpha-fetoprotein (AFP, endoderm). These analyses indicate iPSC lines generated here are indeed pluripotent. Scale bar: 20 µm.
    Tra 1 60 Alexa 488, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 92/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Miltenyi Biotec anti tra 1 60 microbeads
    Verification of the pluripotency of the iPSC lines with the TDP-43 A90V mutation. ( A ) Fluorescence microscopy images of the expression of the pluripotency markers NANOG, OCT4, SSEA4, <t>TRA-1-60,</t> and TRA-1-81 in control (37L25) and patient (36L10) iPSC lines. Scale bar: 20 µm. ( B ) All iPSC lines differentiated into cells of the three germ layers, as shown by expression of desmin (mesoderm), TUJ1 (ectoderm), and alpha-fetoprotein (AFP, endoderm). These analyses indicate iPSC lines generated here are indeed pluripotent. Scale bar: 20 µm.
    Anti Tra 1 60 Microbeads, supplied by Miltenyi Biotec, used in various techniques. Bioz Stars score: 94/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Millipore anti tra 1 60 fitc
    Verification of the pluripotency of the iPSC lines with the TDP-43 A90V mutation. ( A ) Fluorescence microscopy images of the expression of the pluripotency markers NANOG, OCT4, SSEA4, <t>TRA-1-60,</t> and TRA-1-81 in control (37L25) and patient (36L10) iPSC lines. Scale bar: 20 µm. ( B ) All iPSC lines differentiated into cells of the three germ layers, as shown by expression of desmin (mesoderm), TUJ1 (ectoderm), and alpha-fetoprotein (AFP, endoderm). These analyses indicate iPSC lines generated here are indeed pluripotent. Scale bar: 20 µm.
    Anti Tra 1 60 Fitc, supplied by Millipore, used in various techniques. Bioz Stars score: 91/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Becton Dickinson mouse anti tra 1 60
    Verification of the pluripotency of the iPSC lines with the TDP-43 A90V mutation. ( A ) Fluorescence microscopy images of the expression of the pluripotency markers NANOG, OCT4, SSEA4, <t>TRA-1-60,</t> and TRA-1-81 in control (37L25) and patient (36L10) iPSC lines. Scale bar: 20 µm. ( B ) All iPSC lines differentiated into cells of the three germ layers, as shown by expression of desmin (mesoderm), TUJ1 (ectoderm), and alpha-fetoprotein (AFP, endoderm). These analyses indicate iPSC lines generated here are indeed pluripotent. Scale bar: 20 µm.
    Mouse Anti Tra 1 60, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 90/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    hAEC are positive for naïve pluripotent markers. (A) Representative micrographs at 20x from hAEC at P0-P2 immunostained for KLF4 (red) and TFE3 (red); the nuclei were stained with DAPI (blue). (B) Graph shows the percentage of hAEC that express naïve markers. (C) Representative micrographs at 20x from hAEC at P0-P2 immunostained for KLF4 (red) or TFE3 (red) and co-expressing TRA-1-60; the nuclei were stained with DAPI (blue). (D) Graph shows the percentage of cells immunostained for both TRA-1-60 and naïve markers. Results are expressed as percentages of means ± S.E.M. from 9 fields counted in duplicate from three independent experiments. Scale bar 50 μm. * p

    Journal: PLoS ONE

    Article Title: Markers of Pluripotency in Human Amniotic Epithelial Cells and Their Differentiation to Progenitor of Cortical Neurons

    doi: 10.1371/journal.pone.0146082

    Figure Lengend Snippet: hAEC are positive for naïve pluripotent markers. (A) Representative micrographs at 20x from hAEC at P0-P2 immunostained for KLF4 (red) and TFE3 (red); the nuclei were stained with DAPI (blue). (B) Graph shows the percentage of hAEC that express naïve markers. (C) Representative micrographs at 20x from hAEC at P0-P2 immunostained for KLF4 (red) or TFE3 (red) and co-expressing TRA-1-60; the nuclei were stained with DAPI (blue). (D) Graph shows the percentage of cells immunostained for both TRA-1-60 and naïve markers. Results are expressed as percentages of means ± S.E.M. from 9 fields counted in duplicate from three independent experiments. Scale bar 50 μm. * p

    Article Snippet: Primary antibodies to evaluate pluripotency were: rabbit anti-OCT4 (1:100, Cat. ab19857, Abcam, San Francisco, CA, USA), rabbit anti-SOX2 (1:150, Cat. ab97959, Abcam), rabbit anti-NANOG (1:500, Cat. 500-P236, Peprotech), rabbit anti-Kruppel-Like Factor (KLF) 4 (1:200, Cat. sc-20691, Santa Cruz Biotechnology, Dallas, TX, USA), rabbit anti-transcription factor binding to immunoglobulin heavy constant mu (IGHM) enhancer 3 (TFE3) (1:100, Cat. ab97667, Abcam), rat anti-SSEA3 (1:50, Cat. ab16286, Abcam), mouse anti-SSEA4 (1:150, Cat. ab16287, Abcam), mouse anti-TRA-1-60 (1:150, Cat. ab16288, Abcam) and mouse anti-E-cadherin (1:50, Cat. 610181, BD Biosciences, San Jose, CA, USA).

    Techniques: Staining, Expressing

    Proliferation of hAEC that display pluripotent markers in vitro . (A) Representative micrographs at 20X from hAEC at different passages (P0-P3) immunostained for OCT4 (red), SOX2 (red), NANOG (red), SSEA3 (green), SSEA4 (green), TRA-1-60 (green) and E-cadherin (green) and also for Ki67 (green or red); the nuclei were stained with DAPI (blue). (B) Graph shows the percentage of hAEC positive for one of the pluripotency factors and Ki67. Results are expressed as percentages of means ± S.E.M. from 9 fields counted in duplicate from five independent experiments. Scale bar 50 μm.

    Journal: PLoS ONE

    Article Title: Markers of Pluripotency in Human Amniotic Epithelial Cells and Their Differentiation to Progenitor of Cortical Neurons

    doi: 10.1371/journal.pone.0146082

    Figure Lengend Snippet: Proliferation of hAEC that display pluripotent markers in vitro . (A) Representative micrographs at 20X from hAEC at different passages (P0-P3) immunostained for OCT4 (red), SOX2 (red), NANOG (red), SSEA3 (green), SSEA4 (green), TRA-1-60 (green) and E-cadherin (green) and also for Ki67 (green or red); the nuclei were stained with DAPI (blue). (B) Graph shows the percentage of hAEC positive for one of the pluripotency factors and Ki67. Results are expressed as percentages of means ± S.E.M. from 9 fields counted in duplicate from five independent experiments. Scale bar 50 μm.

    Article Snippet: Primary antibodies to evaluate pluripotency were: rabbit anti-OCT4 (1:100, Cat. ab19857, Abcam, San Francisco, CA, USA), rabbit anti-SOX2 (1:150, Cat. ab97959, Abcam), rabbit anti-NANOG (1:500, Cat. 500-P236, Peprotech), rabbit anti-Kruppel-Like Factor (KLF) 4 (1:200, Cat. sc-20691, Santa Cruz Biotechnology, Dallas, TX, USA), rabbit anti-transcription factor binding to immunoglobulin heavy constant mu (IGHM) enhancer 3 (TFE3) (1:100, Cat. ab97667, Abcam), rat anti-SSEA3 (1:50, Cat. ab16286, Abcam), mouse anti-SSEA4 (1:150, Cat. ab16287, Abcam), mouse anti-TRA-1-60 (1:150, Cat. ab16288, Abcam) and mouse anti-E-cadherin (1:50, Cat. 610181, BD Biosciences, San Jose, CA, USA).

    Techniques: In Vitro, Staining

    hAEC display the pluripotent stem cell markers. (A) Representative micrographs at 20X from hAEC at different passages (P0-P3) immunostained for OCT4 (red), SOX2 (red), NANOG (red), SSEA3 (green), SSEA4 (green), TRA-1-60 (green) and E-cadherin (green); the nuclei were stained with DAPI (blue). Arrow indicate that the marker was found in the cytoplasm. (B) Graph shows the percentage of hAEC positive for the different pluripotent markers. Results are expressed as percentages of means ± S.E.M. from 9 fields counted in duplicate from five independent experiments. Scale bar 50 μm.

    Journal: PLoS ONE

    Article Title: Markers of Pluripotency in Human Amniotic Epithelial Cells and Their Differentiation to Progenitor of Cortical Neurons

    doi: 10.1371/journal.pone.0146082

    Figure Lengend Snippet: hAEC display the pluripotent stem cell markers. (A) Representative micrographs at 20X from hAEC at different passages (P0-P3) immunostained for OCT4 (red), SOX2 (red), NANOG (red), SSEA3 (green), SSEA4 (green), TRA-1-60 (green) and E-cadherin (green); the nuclei were stained with DAPI (blue). Arrow indicate that the marker was found in the cytoplasm. (B) Graph shows the percentage of hAEC positive for the different pluripotent markers. Results are expressed as percentages of means ± S.E.M. from 9 fields counted in duplicate from five independent experiments. Scale bar 50 μm.

    Article Snippet: Primary antibodies to evaluate pluripotency were: rabbit anti-OCT4 (1:100, Cat. ab19857, Abcam, San Francisco, CA, USA), rabbit anti-SOX2 (1:150, Cat. ab97959, Abcam), rabbit anti-NANOG (1:500, Cat. 500-P236, Peprotech), rabbit anti-Kruppel-Like Factor (KLF) 4 (1:200, Cat. sc-20691, Santa Cruz Biotechnology, Dallas, TX, USA), rabbit anti-transcription factor binding to immunoglobulin heavy constant mu (IGHM) enhancer 3 (TFE3) (1:100, Cat. ab97667, Abcam), rat anti-SSEA3 (1:50, Cat. ab16286, Abcam), mouse anti-SSEA4 (1:150, Cat. ab16287, Abcam), mouse anti-TRA-1-60 (1:150, Cat. ab16288, Abcam) and mouse anti-E-cadherin (1:50, Cat. 610181, BD Biosciences, San Jose, CA, USA).

    Techniques: Staining, Marker

    iPSC pluripotency expression. iPSCs were characterized using standard pluripotency assays. Representative images are shown. (A) iPSCs showed alkaline phosphatase activity. (B) Immunofluorescence staining showed nuclear marker (red) and cell surface (green) pluripotency marker expressions in iPSCs, including NANOG, OCT4, SOX2, and SSEA4, and TRA-1-60. Hoechst nuclear staining in blue. Scale bars 50 μm. (C) Expression of pluripotency-related endogenous genes, OCT4 and Nanog , was detected in derived iPSCs by RT-PCR analysis. hESCs were used as a positive control (Ctrl+), and human fibroblasts (HFs) were used as a negative control (Ctrl−) for endogenous pluripotency gene expression. (D) Some iPSC clones expressed the transgenes (data not shown), whereas fully reprogrammed iPSCs consistently exhibited lentiviral silencing or attenuation (iFLAG and iFLN). HFs at day 5 p.t. were used as a positive control (Ctrl+ HFs D5 p.t.) for transgene expression, and HFs were used as a negative control (Ctrl− HFs). GAPDH was used as an internal control.

    Journal: Frontiers in Cellular Neuroscience

    Article Title: A Three-Dimensional Alzheimer’s Disease Cell Culture Model Using iPSC-Derived Neurons Carrying A246E Mutation in PSEN1

    doi: 10.3389/fncel.2020.00151

    Figure Lengend Snippet: iPSC pluripotency expression. iPSCs were characterized using standard pluripotency assays. Representative images are shown. (A) iPSCs showed alkaline phosphatase activity. (B) Immunofluorescence staining showed nuclear marker (red) and cell surface (green) pluripotency marker expressions in iPSCs, including NANOG, OCT4, SOX2, and SSEA4, and TRA-1-60. Hoechst nuclear staining in blue. Scale bars 50 μm. (C) Expression of pluripotency-related endogenous genes, OCT4 and Nanog , was detected in derived iPSCs by RT-PCR analysis. hESCs were used as a positive control (Ctrl+), and human fibroblasts (HFs) were used as a negative control (Ctrl−) for endogenous pluripotency gene expression. (D) Some iPSC clones expressed the transgenes (data not shown), whereas fully reprogrammed iPSCs consistently exhibited lentiviral silencing or attenuation (iFLAG and iFLN). HFs at day 5 p.t. were used as a positive control (Ctrl+ HFs D5 p.t.) for transgene expression, and HFs were used as a negative control (Ctrl− HFs). GAPDH was used as an internal control.

    Article Snippet: The following primary antibodies were used: 1:2,000 SOX2 (Abcam Cat# ab97959, RRID: AB_2341193 ), 1:400 Nanog (Cell Signaling Technology Cat# 4903, RRID: AB_10559205 ), 1:1,000 OCT4 (Abcam Cat# ab19857, RRID: AB_445175 ), 1:1,000 TRA-1-60 (Abcam Cat# ab16288, RRID: AB_778563 ), 1:1,000 SSEA4 (Abcam Cat# ab16287, RRID: AB_778073 ), 1:500 Nestin (Abcam Cat# ab18102, RRID: AB_444246 ), 1:500 MAP2 (Abcam Cat# ab11267, RRID: AB_297885 ), 1:1,000 Tuj-1 (Abcam Cat# ab215037), and 1:500 Aβ D54D2 antibody (Cell Signaling Technology Cat# 8243, RRID: AB_2797642 ).

    Techniques: Expressing, Activity Assay, Immunofluorescence, Staining, Marker, Derivative Assay, Reverse Transcription Polymerase Chain Reaction, Positive Control, Negative Control, Clone Assay

    Pluripotency markers expression of bioreactor-expanded hiPSCs. Flow cytometry revealed that the majority of cells expressed pluripotency-associated surface marker TRA 1-60  (A;  isotype control shown in gray ) . Quantitative real time analysis for transcription

    Journal: Tissue Engineering. Part C, Methods

    Article Title: Suspension Culture of Human Pluripotent Stem Cells in Controlled, Stirred Bioreactors

    doi: 10.1089/ten.tec.2011.0717

    Figure Lengend Snippet: Pluripotency markers expression of bioreactor-expanded hiPSCs. Flow cytometry revealed that the majority of cells expressed pluripotency-associated surface marker TRA 1-60 (A; isotype control shown in gray ) . Quantitative real time analysis for transcription

    Article Snippet: Single cell suspensions were prepared and aliquots were incubated with a primary antibody against TRA 1-60 (1:100; mouse IgM, Abcam) and corresponding isotype control for 30 min at 4°C.

    Techniques: Expressing, Flow Cytometry, Cytometry, Marker

    Characterization of iPS cells from clonally expanded MSCs and HDF. A , immunocytochemistry of SSEA-3, SSEA-4, TRA-1–60, TRA-1–81, OCT3/4, and NANOG for iPS cells. Scale bars = 100 μm. B , DNA methylation states of the OCT3/4 and

    Journal: The Journal of Biological Chemistry

    Article Title: Induction of Pluripotent Stem Cells from Human Third Molar Mesenchymal Stromal Cells *

    doi: 10.1074/jbc.M109.055889

    Figure Lengend Snippet: Characterization of iPS cells from clonally expanded MSCs and HDF. A , immunocytochemistry of SSEA-3, SSEA-4, TRA-1–60, TRA-1–81, OCT3/4, and NANOG for iPS cells. Scale bars = 100 μm. B , DNA methylation states of the OCT3/4 and

    Article Snippet: Primary antibodies included stage-specific embryonic antigen (SSEA)-3 (1:100, MAB4303, Millipore), SSEA-4 (1:100, MAB4304, Millipore), tumor-related antigen (TRA)-1–60 (1:100, ab16288-200, Abcam), TRA-1–81 (1:100, ab16289-200, Abcam), OCT4 (1:100, ab19857-100, Abcam), NANOG (1:50, ab21624, Abcam), SOX17 (1:200, AF1924, R & D Systems), α-fetoprotein (1:200, MAB1368, R & D Systems), desmin (1:200, RB-9014, Lab Vision), α-smooth muscle actin (prediluted, N1584, Dako), βIII-tubulin (1:200, CBL412, Millipore), and tyrosine hydroxylase (1:200, AB152, Millipore).

    Techniques: Immunocytochemistry, DNA Methylation Assay

    Hsa-let-7c overexpression alters human induced neuronal (iN) cell morphology. (A) induced pluripotent stem (iPS) cells stain positive for pluripotent markers OCT4 and Tra-1-60. (B) iN cell induction protocol for converting human iPS cells into functional glutamatergic neurons. (C) Immunocytochemical characterization of excitatory iN cells. iN cells transduced with Ngn2 stain positive for the neuronal lineage marker TUJ1, mature neuronal marker MAP2, and presynaptic marker synapsin; Ngn2-iN cells are also positive for the excitatory synaptic marker, vGlut1, and negative for the inhibitory marker vGAT. (D) Design for the let-7c overexpression construct. The H1 promoter and a downstream short hairpin RNA (shRNA) containing the mature let-7c sequence were cloned into the FG12 lentiviral backbone. (E) HEK293T cells were transfected with either FG12 or FG12-let-7c and assayed via qPCR to assess let-7c overexpression. (F–H) The FG12-let-7c construct was packaged into lentivirus and used to infect human iN cells according to (B) . Infected cells express GFP (F) and AG2U cells show a significant reduction in soma size (G) , as well as primary dendrite outgrowth (H) . Statistical tests were performed using two-tailed Student’s t -test, where * p

    Journal: Frontiers in Synaptic Neuroscience

    Article Title: hsa-let-7c miRNA Regulates Synaptic and Neuronal Function in Human Neurons

    doi: 10.3389/fnsyn.2018.00019

    Figure Lengend Snippet: Hsa-let-7c overexpression alters human induced neuronal (iN) cell morphology. (A) induced pluripotent stem (iPS) cells stain positive for pluripotent markers OCT4 and Tra-1-60. (B) iN cell induction protocol for converting human iPS cells into functional glutamatergic neurons. (C) Immunocytochemical characterization of excitatory iN cells. iN cells transduced with Ngn2 stain positive for the neuronal lineage marker TUJ1, mature neuronal marker MAP2, and presynaptic marker synapsin; Ngn2-iN cells are also positive for the excitatory synaptic marker, vGlut1, and negative for the inhibitory marker vGAT. (D) Design for the let-7c overexpression construct. The H1 promoter and a downstream short hairpin RNA (shRNA) containing the mature let-7c sequence were cloned into the FG12 lentiviral backbone. (E) HEK293T cells were transfected with either FG12 or FG12-let-7c and assayed via qPCR to assess let-7c overexpression. (F–H) The FG12-let-7c construct was packaged into lentivirus and used to infect human iN cells according to (B) . Infected cells express GFP (F) and AG2U cells show a significant reduction in soma size (G) , as well as primary dendrite outgrowth (H) . Statistical tests were performed using two-tailed Student’s t -test, where * p

    Article Snippet: The following antibodies were used for our analysis: mouse anti-MAP2 (Sigma, 1:1000), rabbit anti-MAP2 (Sigma, 1:1000), rabbit anti-synapsin (E028, 1:3,000), mouse anti-vGlut1 (Synaptic Systems, 1:200), mouse anti-vGAT (Synaptic Systems, 1:200), rabbit anti-TUJ1 (Synaptic Systems, 1:200), anti-TRA-1-60 (Millipore, 1:1000), anti-Oct4 (Millipore, 1:1000) and anti-MeCP2 (Neuromab, 1:500).

    Techniques: Over Expression, Staining, Functional Assay, Transduction, Marker, Construct, shRNA, Sequencing, Clone Assay, Transfection, Real-time Polymerase Chain Reaction, Infection, Two Tailed Test

    Immunostaining. Fourteen days after embryoid body (EB) formation, cells were subjected to immunostaining with Oct3/4, Nanog, SSEA-4 and TRA-1-60 antibodies. Cells cultured without Activin A were negative for all of the antibodies, while cells were positive

    Journal: Experimental and Therapeutic Medicine

    Article Title: Activin A maintains pluripotency markers and proliferative potential of human induced pluripotent stem cells

    doi: 10.3892/etm.2011.219

    Figure Lengend Snippet: Immunostaining. Fourteen days after embryoid body (EB) formation, cells were subjected to immunostaining with Oct3/4, Nanog, SSEA-4 and TRA-1-60 antibodies. Cells cultured without Activin A were negative for all of the antibodies, while cells were positive

    Article Snippet: Diluted (1:500) anti-Oct3/4, anti-Nanog, anti-SSEA-4 and anti-TRA-1-60 antibodies (all from Nihon Millipore K.K., Tokyo, Japan) were incubated in wash buffer overnight at 4°C.

    Techniques: Immunostaining, Cell Culture

    hPSCs cultured in the defined, xeno-free EOD hPSC medium retain their stem cell characteristics. Immunodetection of pluripotency antigens in hPSC lines cultivated for 40 passages in the defined, xeno-free L7 hPSC medium with EOD medium changes. Detection of Oct-3/4 and Nanog are shown (red) along with SSEA-4 and TRA-1-60 (green). Individual cell nuclei were visualized using DAPI (blue). Scale bar: 200 μm.

    Journal: PLoS ONE

    Article Title: A Newly Defined and Xeno-Free Culture Medium Supports Every-Other-Day Medium Replacement in the Generation and Long-Term Cultivation of Human Pluripotent Stem Cells

    doi: 10.1371/journal.pone.0161229

    Figure Lengend Snippet: hPSCs cultured in the defined, xeno-free EOD hPSC medium retain their stem cell characteristics. Immunodetection of pluripotency antigens in hPSC lines cultivated for 40 passages in the defined, xeno-free L7 hPSC medium with EOD medium changes. Detection of Oct-3/4 and Nanog are shown (red) along with SSEA-4 and TRA-1-60 (green). Individual cell nuclei were visualized using DAPI (blue). Scale bar: 200 μm.

    Article Snippet: Extracellular antigens were detected on unfixed cells stained with PE-conjugated antigen-specific antibodies and respective isotypes using the manufacturer’s recommended concentration: anti-TRA-1-60 (BD Biosciences, 560193), anti-TRA-1-81 (BD Biosciences, 560161), mouse IgG3 isotype (BD Biosciences, 556659); anti-SSEA4 (BD Biosciences, 560128) and mouse IgM isotype (BD Biosciences, 555584).

    Techniques: Cell Culture, Immunodetection

    Expression of pluripotency markers in hPSCs cultured for 40 passages using the defined, xeno-free EOD hPSC medium. Three hPSC lines (two replicates per line) were evaluated for their ability to sustain the expression of markers of pluripotency at passage 10, passage 25 and passage 40. Each hPSC line was independently analyzed by flow cytometry for the expression of SSEA-4, TRA-1-60 and TRA-1-81. Results within each passage were combined and the mean value of marker expression determined. Bars represent mean percentage values. Error bars represent the standard deviation of the mean for each passage.

    Journal: PLoS ONE

    Article Title: A Newly Defined and Xeno-Free Culture Medium Supports Every-Other-Day Medium Replacement in the Generation and Long-Term Cultivation of Human Pluripotent Stem Cells

    doi: 10.1371/journal.pone.0161229

    Figure Lengend Snippet: Expression of pluripotency markers in hPSCs cultured for 40 passages using the defined, xeno-free EOD hPSC medium. Three hPSC lines (two replicates per line) were evaluated for their ability to sustain the expression of markers of pluripotency at passage 10, passage 25 and passage 40. Each hPSC line was independently analyzed by flow cytometry for the expression of SSEA-4, TRA-1-60 and TRA-1-81. Results within each passage were combined and the mean value of marker expression determined. Bars represent mean percentage values. Error bars represent the standard deviation of the mean for each passage.

    Article Snippet: Extracellular antigens were detected on unfixed cells stained with PE-conjugated antigen-specific antibodies and respective isotypes using the manufacturer’s recommended concentration: anti-TRA-1-60 (BD Biosciences, 560193), anti-TRA-1-81 (BD Biosciences, 560161), mouse IgG3 isotype (BD Biosciences, 556659); anti-SSEA4 (BD Biosciences, 560128) and mouse IgM isotype (BD Biosciences, 555584).

    Techniques: Expressing, Cell Culture, Flow Cytometry, Cytometry, Marker, Standard Deviation

    In vitro characterization of the pluripotent state of EiPS cells. ( a ). Nanog, SSEA1, SSEA4, TRA-1-60 and TRA-1-81 detected by fluorescent immunohistochemistry. Yellow scale bar 10 μm, white scalebar 20 μm. ( b ). Detection of gene expression of key pluripotency markers using RT-PCR primers specific for equine endogenous genes

    Journal: Stem Cell Reviews

    Article Title: Induced Pluripotent Stem Cell Lines Derived from Equine Fibroblasts

    doi: 10.1007/s12015-011-9239-5

    Figure Lengend Snippet: In vitro characterization of the pluripotent state of EiPS cells. ( a ). Nanog, SSEA1, SSEA4, TRA-1-60 and TRA-1-81 detected by fluorescent immunohistochemistry. Yellow scale bar 10 μm, white scalebar 20 μm. ( b ). Detection of gene expression of key pluripotency markers using RT-PCR primers specific for equine endogenous genes

    Article Snippet: The cells were then incubated overnight at 4°C with the following primary anti-mouse or anti-human antibodies; (i ) anti-Nanog (Reprocell #RCAB0002P-F), (ii ) anti-Oct4 (Santa Cruz #sc-5279), (iii ) anti-SSEA1 (Stemgent #09-0005), (vi) anti-SSEA4 (Stemgent #09-0006), (v ) anti-TRA-1-60 (Stemgent #09-0009), (vi ) anti-TRA-1-81 (Stemgent #09-0011).

    Techniques: In Vitro, Immunohistochemistry, Expressing, Reverse Transcription Polymerase Chain Reaction

    XPC increases somatic reprogramming fidelity and iPSC self-renewal capacity. ( A . ( B ) Flow cytometry analysis of control and XPC complex (XPC–RAD23B–CETN2) gain-of-function cell populations 24 d post-induction for the late stage human iPSC marker TRA-1-60. ( C ) The average number of colonies obtained in the reprogramming experiment depicted in B . ( D ) iPSCs derived from control or XPC–RAD23B–CETN2-overexpressing HDFs were challenged with single-cell dissociation and allowed to recover for 3–4 d before staining with alkaline phosphatase (AP). The average number of AP + colonies formed per 2.5 × 10 4 single cells plated. Error bars depict the standard deviation. n = 3. (*) P

    Journal: Genes & Development

    Article Title: Regulation of DNA demethylation by the XPC DNA repair complex in somatic and pluripotent stem cells

    doi: 10.1101/gad.295741.116

    Figure Lengend Snippet: XPC increases somatic reprogramming fidelity and iPSC self-renewal capacity. ( A . ( B ) Flow cytometry analysis of control and XPC complex (XPC–RAD23B–CETN2) gain-of-function cell populations 24 d post-induction for the late stage human iPSC marker TRA-1-60. ( C ) The average number of colonies obtained in the reprogramming experiment depicted in B . ( D ) iPSCs derived from control or XPC–RAD23B–CETN2-overexpressing HDFs were challenged with single-cell dissociation and allowed to recover for 3–4 d before staining with alkaline phosphatase (AP). The average number of AP + colonies formed per 2.5 × 10 4 single cells plated. Error bars depict the standard deviation. n = 3. (*) P

    Article Snippet: The commercial antibodies used were as follows: anti-ACTB (A2228) from Sigma-Aldrich, anti-XPC (A301-122A) and anti-RAD23B (A302-306A) from Bethyl Laboratories, anti-CETN2 (15977-1-AP) from ProteinTech, anti-5mC (33D3) from Diagenode, and anti-SSEA4 (clone MC-813-70) and anti-TRA-1-60 (clone TRA-1-60-R) from Biolegends.

    Techniques: Flow Cytometry, Cytometry, Marker, Derivative Assay, Staining, Standard Deviation

    Characterization of the two types of piPSCs. (A)The colonies of mpiPSCs were round and three-dimensional (bottom). The colonies of hpiPSCs were large and flat (top). Bar scale = 500 μm. The hpiPSCs and mpiPSCs were both positive for alkaline phosphatase (AP). Bar scale = 200 μm. (B) Immunocytochemical staining of Oct4, Sox2, SSEA-1 and SSEA-4 in hpiPSCs and mpiPSCs. Bar scale = 200 μm. (C) The surface marker Tra-1-81 and Tra-1-60 were positive in hpiPSCs but not detected in mpiPSCs. Bar scale = 200 μm. (D) X chromosome activation state of hpiPSCs and mpiPSCs after immunostaining for H3K27me3. Positive signals of histone H3K27 me3 spots were observed in hpiPSCs but not in mpiPSCs. Bar scale = 200 μm. The white arrowheads indicate the H3K27me3 positive spot in the cell. (E) Immunocytochemical assay of 3-germ-layer cells in EBs derived from both types of piPSCs, the markers include β Ⅲ-Tubulin (ectoderm), α-SMA (mesoderm) and Sox17 (endoderm). Scale bar = 200 μm. (F) Hematoxylin and eosin staining of teratoma sections of piPSCs. Left: blood vessel of endothelium (ectoderm); middle: muscle (mesoderm); right: gut-like epithelium (endoderm). Scale bars = 200 μm. (G) Karyotype analysis of piPSCs.

    Journal: PLoS ONE

    Article Title: Pluripotent and Metabolic Features of Two Types of Porcine iPSCs Derived from Defined Mouse and Human ES Cell Culture Conditions

    doi: 10.1371/journal.pone.0124562

    Figure Lengend Snippet: Characterization of the two types of piPSCs. (A)The colonies of mpiPSCs were round and three-dimensional (bottom). The colonies of hpiPSCs were large and flat (top). Bar scale = 500 μm. The hpiPSCs and mpiPSCs were both positive for alkaline phosphatase (AP). Bar scale = 200 μm. (B) Immunocytochemical staining of Oct4, Sox2, SSEA-1 and SSEA-4 in hpiPSCs and mpiPSCs. Bar scale = 200 μm. (C) The surface marker Tra-1-81 and Tra-1-60 were positive in hpiPSCs but not detected in mpiPSCs. Bar scale = 200 μm. (D) X chromosome activation state of hpiPSCs and mpiPSCs after immunostaining for H3K27me3. Positive signals of histone H3K27 me3 spots were observed in hpiPSCs but not in mpiPSCs. Bar scale = 200 μm. The white arrowheads indicate the H3K27me3 positive spot in the cell. (E) Immunocytochemical assay of 3-germ-layer cells in EBs derived from both types of piPSCs, the markers include β Ⅲ-Tubulin (ectoderm), α-SMA (mesoderm) and Sox17 (endoderm). Scale bar = 200 μm. (F) Hematoxylin and eosin staining of teratoma sections of piPSCs. Left: blood vessel of endothelium (ectoderm); middle: muscle (mesoderm); right: gut-like epithelium (endoderm). Scale bars = 200 μm. (G) Karyotype analysis of piPSCs.

    Article Snippet: The following primary antibodies were used: anti-Oct4 (Santa cruz, sc-5297), anti-Sox2 (Abcam, ab97959), anti-SSEA-1 (Cell Signaling Technology, 4744S), anti-SSEA-4 (Cell Signaling Technology, 4755P), anti-Tra-1-81 (Cell Signaling Technology, 4745P), anti-Tra-1-60 (Cell Signaling Technology, 4746P) and anti-H3K27me3 (Abcam, ab6002).

    Techniques: Staining, Marker, Activation Assay, Immunostaining, Derivative Assay

    Confirmation of pluripotent markers in cells and tissue samples. ( a ) Relative gene expression of OCT4. ( b ) Relative gene expression of LIN28 in cells and pellets. ( c ) Fluorescence image of iPSC-derived teratoma tissues stained with the TRA-1-60 and c-Myc antibody. Fluorescence image of ( d ) mcMock, ( e ) mcBMP2, ( f ) mcTGFβ3, and ( g ) mcBOTH. Images show the 200× magnification image. Scale bar represents 200 μm. mcMock: minicircle mock vector-transfected outgrowth cells; mcBMP2: minicircle BMP2-encoding vector-transfected outgrowth cells; mcTGFβ3: minicircle TGFβ3-encoding vector-transfected outgrowth cells; mcBOTH: 1:1 mixture of mcBMP2- and mc TGFβ3-transfected outgrowth cells.

    Journal: Cells

    Article Title: Chondrogenic Differentiation from Induced Pluripotent Stem Cells Using Non-Viral Minicircle Vectors

    doi: 10.3390/cells9030582

    Figure Lengend Snippet: Confirmation of pluripotent markers in cells and tissue samples. ( a ) Relative gene expression of OCT4. ( b ) Relative gene expression of LIN28 in cells and pellets. ( c ) Fluorescence image of iPSC-derived teratoma tissues stained with the TRA-1-60 and c-Myc antibody. Fluorescence image of ( d ) mcMock, ( e ) mcBMP2, ( f ) mcTGFβ3, and ( g ) mcBOTH. Images show the 200× magnification image. Scale bar represents 200 μm. mcMock: minicircle mock vector-transfected outgrowth cells; mcBMP2: minicircle BMP2-encoding vector-transfected outgrowth cells; mcTGFβ3: minicircle TGFβ3-encoding vector-transfected outgrowth cells; mcBOTH: 1:1 mixture of mcBMP2- and mc TGFβ3-transfected outgrowth cells.

    Article Snippet: For the confirmation of pluripotency, the anti-c-Myc (1:250; ab32072, Abcam), and anti-TRA-1-60 (1:200; MAB4360, Merck Millipore, Burlington, MA, USA) antibodies were used.

    Techniques: Expressing, Fluorescence, Derivative Assay, Staining, Plasmid Preparation, Transfection

    Undifferentiated ORMES6 ESCs express embryonic cell markers. (A) SSEA 4 (green); (B) Oct4 (red); (C) glycoprotein TRA-1-60 (green)/ Oct4 (red) and (D) glycoprotein TRA-1-81 (green)/Oct4 (red). The nuclei were stained with DAPI (blue). The undifferentiated ORMES6 ESCs expressed SSEA 4, Oct4, TRA-1-60 and TRA-1-81 antigens. Cells only stained with DAPI were MEF feeder cells. Scale bar: 40 μm.

    Journal: Differentiation; research in biological diversity

    Article Title: Differentiation of nonhuman primate embryonic stem cells along neural lineages

    doi: 10.1016/j.diff.2008.10.014

    Figure Lengend Snippet: Undifferentiated ORMES6 ESCs express embryonic cell markers. (A) SSEA 4 (green); (B) Oct4 (red); (C) glycoprotein TRA-1-60 (green)/ Oct4 (red) and (D) glycoprotein TRA-1-81 (green)/Oct4 (red). The nuclei were stained with DAPI (blue). The undifferentiated ORMES6 ESCs expressed SSEA 4, Oct4, TRA-1-60 and TRA-1-81 antigens. Cells only stained with DAPI were MEF feeder cells. Scale bar: 40 μm.

    Article Snippet: The following primary antibodies were used: anti-neuronal nuclear antigen (NeuN) (IgG, 1:100), anti-tubulin betaIII (IgG, 1:50), anti-nestin (rabbit polyclonal, 1:100), anti-FoxD3 (rabbit polyclonal, 1:50), anti-Pax6 (rabbit polyclonal, 1:100, Covance Research Products, Inc, Denver, PA), anti-microtubule-associated protein (MAP2) (IgG, 1:50), anti-A2B5 (IgM, 1:100), anti-glial fibrillary acidic protein (GFAP) (IgG, 1:100), anti-O4 (IgG, 1:100), anti-galactocerebroside (GalCer) (IgG, 1:100), anti-S100 (rabbit polyclonal, 1:100), anti-SSEA 4 (IgG, 1:66), anti-TRA-1-60 (IgM, 1:66), anti-TRA-1-81(IgM, 1:66), anti-Oct4 (rabbit polyclonal, 1:13.3, Santa Cruz Biotechnology, Santa Cruz, CA), anti-human Ki67 (IgG1, 1:100, DAKO, Carpinteria, CA) and anti-Brachyury (goat polyclonal, 1:13.3, Santa Cruz Biotechnology, Santa Cruz, CA).

    Techniques: Staining

    Verification of the pluripotency of the iPSC lines with the TDP-43 A90V mutation. ( A ) Fluorescence microscopy images of the expression of the pluripotency markers NANOG, OCT4, SSEA4, TRA-1-60, and TRA-1-81 in control (37L25) and patient (36L10) iPSC lines. Scale bar: 20 µm. ( B ) All iPSC lines differentiated into cells of the three germ layers, as shown by expression of desmin (mesoderm), TUJ1 (ectoderm), and alpha-fetoprotein (AFP, endoderm). These analyses indicate iPSC lines generated here are indeed pluripotent. Scale bar: 20 µm.

    Journal: PLoS ONE

    Article Title: Downregulation of MicroRNA-9 in iPSC-Derived Neurons of FTD/ALS Patients with TDP-43 Mutations

    doi: 10.1371/journal.pone.0076055

    Figure Lengend Snippet: Verification of the pluripotency of the iPSC lines with the TDP-43 A90V mutation. ( A ) Fluorescence microscopy images of the expression of the pluripotency markers NANOG, OCT4, SSEA4, TRA-1-60, and TRA-1-81 in control (37L25) and patient (36L10) iPSC lines. Scale bar: 20 µm. ( B ) All iPSC lines differentiated into cells of the three germ layers, as shown by expression of desmin (mesoderm), TUJ1 (ectoderm), and alpha-fetoprotein (AFP, endoderm). These analyses indicate iPSC lines generated here are indeed pluripotent. Scale bar: 20 µm.

    Article Snippet: Cultures were then stained overnight at 4°C with goat anti-NANOG (1∶100, R & D, AF1997), mouse anti-OCT3/4 (1∶100, Santa Cruz Biotechnology, sc-5279), mouse anti-SSEA4 (1∶100, Abcam, ab16287), mouse anti-TRA-1-60 (1∶50, Chemicon, MAB4360), mouse anti-TRA-1-81 (1∶100, Chemicon, MAB4381), rabbit anti-desmin (Thermo Scientific; 1∶100, RB-9014-P0), mouse anti-α-fetoprotein (AFP, R & D Systems; 1∶200, MAB1368), rabbit anti-TDP-43 (1∶200, Proteintech Group, 10782-2-AP), mouse anti-TUJ1 (1∶500, Promega, G712A), rabbit anti-VGLUT1 (1∶500, Synaptic Systems,135303)and rabbit anti-GABA (1∶1000, Sigma, A2052).

    Techniques: Mutagenesis, Fluorescence, Microscopy, Expressing, Generated