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resource source identifier sox2 antibody r&d systems cat#af2018  (R&D Systems)


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    R&D Systems resource source identifier sox2 antibody r&d systems cat#af2018
    Resource Source Identifier Sox2 Antibody R&D Systems Cat#Af2018, supplied by R&D Systems, used in various techniques. Bioz Stars score: 97/100, based on 729 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/resource+source+identifier+antibodies+human+mouse+rat+sox2+antibody+r+d+systems/Human%2FMouse%2FRat+SOX2+Antibody/pm41923624-240-2-8
    Average 97 stars, based on 729 article reviews
    resource source identifier sox2 antibody r&d systems cat#af2018 - by Bioz Stars, 2026-10
    97/100 stars

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    Purification:

    Article Title: Exploring roles of neuroglobin in oxygen homeostasis and neurodevelopment based on human cerebral organoid models.
    Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Human/Mouse/Rat SOX2 Antibody R&D Systems Cat# AF2018, RRID: AB_355110 Anti-PSD95 Antibody Abcam Cat# ab18258, RRID: AB_444362 Purified Mouse Anti-Ki-67 Clone B56 BD Biosciences Cat# 550609, RRID: AB_393778 Anti-NESTIN Antibody Merck Millipore Cat# ABD69, RRID: AB_2744681 Anti-TBR1 Antibody Abcam Cat# ab31940, RRID: AB_2200219 Goat anti-Mouse IgG (H + L) Cross-Adsorbed Secondary Antibody, Alexa Fluor TM 488 Thermo Fisher Scientific Cat# A-11001, RRID: AB_2534069 Goat Anti-Rabbit IgG (H + L) Highly Crossadsorbed Antibody, Alexa Fluor TM 546 Thermo Fisher Scientific Cat# A-11035, RRID: AB_143051 Donkey Anti-Goat IgG (H + L) Antibody, Alexa Fluor TM 647 Thermo Fisher Scientific Cat# A-21082, RRID: AB_141493 Chemicals, peptides, and recombinant proteins Stemflex TM Thermo Fisher Scientific Cat# A3349401 AggreWell TM EB formation medium STEMCELL Technologies Cat# 05893 DMEM/F12 Thermo Fisher Scientific Cat# 12660012 Neurobasal Thermo Fisher Scientific Cat# 21103049 Matrigel Corning Cat# 354277 Polybrene Sigma-Aldrich Cat# H9268 Puromycin Sigma-Aldrich Cat# P8833 Accumax Sigma-Aldrich Cat# A7089 ROCK inhibitor Y-27632 Sigma-Aldrich Cat# SCM075 Glutamax Thermo Fisher Scientific Cat# 35050061 MEM-NEAA Thermo Fisher Scientific Cat#11140050 Heparin Sigma-Aldrich Cat# H0878 Cyclopamine A Sigma-Aldrich Cat# H0878 IWP-2 Sigma-Aldrich Cat# I0536 SAG Sigma-Aldrich Cat# 566660 N2 supplement Thermo Fisher Scientific Cat# 17502048 B27 supplement without vitamin A Thermo Fisher Scientific Cat#12587010 B27 supplement Thermo Fisher Scientific Cat#17504044 2-mercaptoethanol Thermo Fisher Scientific Cat#21985023 Insulin Sigma-Aldrich Cat# I9278 Calcium- and magnesium-free DPBS Thermo Fisher Scientific Cat# 14190144 Cell Recovery Solution Corning Cat# 354253 Neural Tissue Dissociation Kit Miltenyi Biotec Cat# 130-092-628 Calcein AM Thermo Fisher Scientific Cat# C1430 Polyethyleneimine Sigma-Aldrich Cat# P3143 Laminin Sigma-Aldrich Cat# 23017015 Paraformaldehyde Sigma-Aldrich Cat# 158127 Sucrose Sigma-Aldrich Cat# S0389 Gelatin Sigma-Aldrich Cat# G2500 Optimal cutting temperature, OCT compound Leica Biosystems Cat# 3801480 Glycine Sigma-Aldrich Cat# G8898 Triton X-100 Sigma-Aldrich Cat# X100 (Continued on next page) e1 iScience 29, 114997, March 20, 2026 .. Feeder-free human induced pluripotent stem cells (hiPSCs) were obtained from and authenticated by the RIKEN Bioresource Center (Cell No. HPS0076, clone 409B2) and maintained on human embryonic stem cell (hESC)-qualified Matrigel (Corning, NY) coated 35 mm dishes in Stemflex (Thermo Fisher Scientific, Waltham, MA) medium and incubated at 37 ◦ C in a humidified atmosphere containing 5% CO 2 .

    Recombinant:

    Article Title: Exploring roles of neuroglobin in oxygen homeostasis and neurodevelopment based on human cerebral organoid models.
    Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Human/Mouse/Rat SOX2 Antibody R&D Systems Cat# AF2018, RRID: AB_355110 Anti-PSD95 Antibody Abcam Cat# ab18258, RRID: AB_444362 Purified Mouse Anti-Ki-67 Clone B56 BD Biosciences Cat# 550609, RRID: AB_393778 Anti-NESTIN Antibody Merck Millipore Cat# ABD69, RRID: AB_2744681 Anti-TBR1 Antibody Abcam Cat# ab31940, RRID: AB_2200219 Goat anti-Mouse IgG (H + L) Cross-Adsorbed Secondary Antibody, Alexa Fluor TM 488 Thermo Fisher Scientific Cat# A-11001, RRID: AB_2534069 Goat Anti-Rabbit IgG (H + L) Highly Crossadsorbed Antibody, Alexa Fluor TM 546 Thermo Fisher Scientific Cat# A-11035, RRID: AB_143051 Donkey Anti-Goat IgG (H + L) Antibody, Alexa Fluor TM 647 Thermo Fisher Scientific Cat# A-21082, RRID: AB_141493 Chemicals, peptides, and recombinant proteins Stemflex TM Thermo Fisher Scientific Cat# A3349401 AggreWell TM EB formation medium STEMCELL Technologies Cat# 05893 DMEM/F12 Thermo Fisher Scientific Cat# 12660012 Neurobasal Thermo Fisher Scientific Cat# 21103049 Matrigel Corning Cat# 354277 Polybrene Sigma-Aldrich Cat# H9268 Puromycin Sigma-Aldrich Cat# P8833 Accumax Sigma-Aldrich Cat# A7089 ROCK inhibitor Y-27632 Sigma-Aldrich Cat# SCM075 Glutamax Thermo Fisher Scientific Cat# 35050061 MEM-NEAA Thermo Fisher Scientific Cat#11140050 Heparin Sigma-Aldrich Cat# H0878 Cyclopamine A Sigma-Aldrich Cat# H0878 IWP-2 Sigma-Aldrich Cat# I0536 SAG Sigma-Aldrich Cat# 566660 N2 supplement Thermo Fisher Scientific Cat# 17502048 B27 supplement without vitamin A Thermo Fisher Scientific Cat#12587010 B27 supplement Thermo Fisher Scientific Cat#17504044 2-mercaptoethanol Thermo Fisher Scientific Cat#21985023 Insulin Sigma-Aldrich Cat# I9278 Calcium- and magnesium-free DPBS Thermo Fisher Scientific Cat# 14190144 Cell Recovery Solution Corning Cat# 354253 Neural Tissue Dissociation Kit Miltenyi Biotec Cat# 130-092-628 Calcein AM Thermo Fisher Scientific Cat# C1430 Polyethyleneimine Sigma-Aldrich Cat# P3143 Laminin Sigma-Aldrich Cat# 23017015 Paraformaldehyde Sigma-Aldrich Cat# 158127 Sucrose Sigma-Aldrich Cat# S0389 Gelatin Sigma-Aldrich Cat# G2500 Optimal cutting temperature, OCT compound Leica Biosystems Cat# 3801480 Glycine Sigma-Aldrich Cat# G8898 Triton X-100 Sigma-Aldrich Cat# X100 (Continued on next page) e1 iScience 29, 114997, March 20, 2026 .. Feeder-free human induced pluripotent stem cells (hiPSCs) were obtained from and authenticated by the RIKEN Bioresource Center (Cell No. HPS0076, clone 409B2) and maintained on human embryonic stem cell (hESC)-qualified Matrigel (Corning, NY) coated 35 mm dishes in Stemflex (Thermo Fisher Scientific, Waltham, MA) medium and incubated at 37 ◦ C in a humidified atmosphere containing 5% CO 2 .

    Cell Recovery:

    Article Title: Exploring roles of neuroglobin in oxygen homeostasis and neurodevelopment based on human cerebral organoid models.
    Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Human/Mouse/Rat SOX2 Antibody R&D Systems Cat# AF2018, RRID: AB_355110 Anti-PSD95 Antibody Abcam Cat# ab18258, RRID: AB_444362 Purified Mouse Anti-Ki-67 Clone B56 BD Biosciences Cat# 550609, RRID: AB_393778 Anti-NESTIN Antibody Merck Millipore Cat# ABD69, RRID: AB_2744681 Anti-TBR1 Antibody Abcam Cat# ab31940, RRID: AB_2200219 Goat anti-Mouse IgG (H + L) Cross-Adsorbed Secondary Antibody, Alexa Fluor TM 488 Thermo Fisher Scientific Cat# A-11001, RRID: AB_2534069 Goat Anti-Rabbit IgG (H + L) Highly Crossadsorbed Antibody, Alexa Fluor TM 546 Thermo Fisher Scientific Cat# A-11035, RRID: AB_143051 Donkey Anti-Goat IgG (H + L) Antibody, Alexa Fluor TM 647 Thermo Fisher Scientific Cat# A-21082, RRID: AB_141493 Chemicals, peptides, and recombinant proteins Stemflex TM Thermo Fisher Scientific Cat# A3349401 AggreWell TM EB formation medium STEMCELL Technologies Cat# 05893 DMEM/F12 Thermo Fisher Scientific Cat# 12660012 Neurobasal Thermo Fisher Scientific Cat# 21103049 Matrigel Corning Cat# 354277 Polybrene Sigma-Aldrich Cat# H9268 Puromycin Sigma-Aldrich Cat# P8833 Accumax Sigma-Aldrich Cat# A7089 ROCK inhibitor Y-27632 Sigma-Aldrich Cat# SCM075 Glutamax Thermo Fisher Scientific Cat# 35050061 MEM-NEAA Thermo Fisher Scientific Cat#11140050 Heparin Sigma-Aldrich Cat# H0878 Cyclopamine A Sigma-Aldrich Cat# H0878 IWP-2 Sigma-Aldrich Cat# I0536 SAG Sigma-Aldrich Cat# 566660 N2 supplement Thermo Fisher Scientific Cat# 17502048 B27 supplement without vitamin A Thermo Fisher Scientific Cat#12587010 B27 supplement Thermo Fisher Scientific Cat#17504044 2-mercaptoethanol Thermo Fisher Scientific Cat#21985023 Insulin Sigma-Aldrich Cat# I9278 Calcium- and magnesium-free DPBS Thermo Fisher Scientific Cat# 14190144 Cell Recovery Solution Corning Cat# 354253 Neural Tissue Dissociation Kit Miltenyi Biotec Cat# 130-092-628 Calcein AM Thermo Fisher Scientific Cat# C1430 Polyethyleneimine Sigma-Aldrich Cat# P3143 Laminin Sigma-Aldrich Cat# 23017015 Paraformaldehyde Sigma-Aldrich Cat# 158127 Sucrose Sigma-Aldrich Cat# S0389 Gelatin Sigma-Aldrich Cat# G2500 Optimal cutting temperature, OCT compound Leica Biosystems Cat# 3801480 Glycine Sigma-Aldrich Cat# G8898 Triton X-100 Sigma-Aldrich Cat# X100 (Continued on next page) e1 iScience 29, 114997, March 20, 2026 .. Feeder-free human induced pluripotent stem cells (hiPSCs) were obtained from and authenticated by the RIKEN Bioresource Center (Cell No. HPS0076, clone 409B2) and maintained on human embryonic stem cell (hESC)-qualified Matrigel (Corning, NY) coated 35 mm dishes in Stemflex (Thermo Fisher Scientific, Waltham, MA) medium and incubated at 37 ◦ C in a humidified atmosphere containing 5% CO 2 .



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    Figure 1. CELF2 pathogenic variants disrupt nucleocytoplasmic localization and cortical development (A) Selected MR images. Sagittal T1-weighted (top row) and coronal T2-weighted (bottom row) images of patient 1 (i and i0), patient 2 (ii and ii0), patient 3 (iii and iii0), and patient 4 (iv and iv0) are shown. The images were obtained at age 25 months for patient 1, 16 months for patient 2, 25 months for patient 3, and 41 months for patient 4. All patients have simplified gyral pattern, which is mild in patients 1, 3, and 4, and is moderate in patient 2. The corpus callosum is healthy in patient 1, normally developed but diffusely thinned in patient 2, healthy in patient 3, and mildly hypoplastic in patient 4. Patients 1 and 3 have normally developed anterior commissure (arrows in i, ii0, iii, and iii0), but patients 2 and 4 have an absence of the anterior commissure. Patient 2 has moderate ventriculomegaly. (B) The schematic showing the location of the identified variants in the coding exons and the corresponding parts of the CELF2 protein with three RNA-recognition motifs (RRM1–3) (Ladd and Cooper, 2004). The affected residues are within a known nuclear-localization signal (NLS) in RRM3. (C) Boxplots showing CELFs expression in human cortical NPCs (PAX6+ and <t>SOX2+)</t> (single-cell RNA-seq dataset from Pollen et al., 2015). (D) Temporal quantitative real-time PCR analysis of the expression of CELF1 and CELF2 in the developing mouse cortices at the indicated time points. n = 3 samples at each time point. (E) Confocal images of HEK293 cells expressing EGFP-CELF2 (green) that carry different pathogenic variants and counterstained for Hoechst 33258 (blue). ‘‘N’’ denotes the nucleus. Dashed white lines denote nuclei (blue). The intensity of CELF2 immunosignals across individual cells is shown at the bottom with the nucleus region labeled by the blue bar. Scale bar, 10 mm. (F) RT-PCR analysis of exon-skipping events for PPP1R mRNA and TRAF3 mRNA in HEK293 cells transfected with empty vector (control), wild-type (WT), or mutant CELF2. (G) Quantifications of the percentage of transcripts with the skipped exon. One-way ANOVA with Tukey’s test. n = 3 experiments. (H) The quantitative real-time PCR enrichment analysis of GAPDH mRNA and MCL mRNA by RIP with control IgG or an anti-EGFP antibody in HEK293 cells transfected with EGFP-tagged CELF2-NES or mutant CELF2 (normalized to the total RNA input). n = 3 experiments. Data are presented as means ± SEM. Each symbol represents one experiment.
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    Figure 2. Attenuation of Hippo Signaling Promotes Nuclear Accumulation of YAP in Epiblast (A) Distribution of YAP and <t>SOX2</t> proteins in early, mid and late blastocyst stage embryos. Nuclear YAP signals gradually increase in the forming epiblast. Dashed lines indicate SOX2-positive ICM or epiblast cells. (A0) Enlargement of (A). (B) Quantification of nuclear YAP signals.
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    Figure 1. CELF2 pathogenic variants disrupt nucleocytoplasmic localization and cortical development (A) Selected MR images. Sagittal T1-weighted (top row) and coronal T2-weighted (bottom row) images of patient 1 (i and i0), patient 2 (ii and ii0), patient 3 (iii and iii0), and patient 4 (iv and iv0) are shown. The images were obtained at age 25 months for patient 1, 16 months for patient 2, 25 months for patient 3, and 41 months for patient 4. All patients have simplified gyral pattern, which is mild in patients 1, 3, and 4, and is moderate in patient 2. The corpus callosum is healthy in patient 1, normally developed but diffusely thinned in patient 2, healthy in patient 3, and mildly hypoplastic in patient 4. Patients 1 and 3 have normally developed anterior commissure (arrows in i, ii0, iii, and iii0), but patients 2 and 4 have an absence of the anterior commissure. Patient 2 has moderate ventriculomegaly. (B) The schematic showing the location of the identified variants in the coding exons and the corresponding parts of the CELF2 protein with three RNA-recognition motifs (RRM1–3) (Ladd and Cooper, 2004). The affected residues are within a known nuclear-localization signal (NLS) in RRM3. (C) Boxplots showing CELFs expression in human cortical NPCs (PAX6+ and SOX2+) (single-cell RNA-seq dataset from Pollen et al., 2015). (D) Temporal quantitative real-time PCR analysis of the expression of CELF1 and CELF2 in the developing mouse cortices at the indicated time points. n = 3 samples at each time point. (E) Confocal images of HEK293 cells expressing EGFP-CELF2 (green) that carry different pathogenic variants and counterstained for Hoechst 33258 (blue). ‘‘N’’ denotes the nucleus. Dashed white lines denote nuclei (blue). The intensity of CELF2 immunosignals across individual cells is shown at the bottom with the nucleus region labeled by the blue bar. Scale bar, 10 mm. (F) RT-PCR analysis of exon-skipping events for PPP1R mRNA and TRAF3 mRNA in HEK293 cells transfected with empty vector (control), wild-type (WT), or mutant CELF2. (G) Quantifications of the percentage of transcripts with the skipped exon. One-way ANOVA with Tukey’s test. n = 3 experiments. (H) The quantitative real-time PCR enrichment analysis of GAPDH mRNA and MCL mRNA by RIP with control IgG or an anti-EGFP antibody in HEK293 cells transfected with EGFP-tagged CELF2-NES or mutant CELF2 (normalized to the total RNA input). n = 3 experiments. Data are presented as means ± SEM. Each symbol represents one experiment.

    Journal: Cell reports

    Article Title: Nucleocytoplasmic transport of the RNA-binding protein CELF2 regulates neural stem cell fates.

    doi: 10.1016/j.celrep.2021.109226

    Figure Lengend Snippet: Figure 1. CELF2 pathogenic variants disrupt nucleocytoplasmic localization and cortical development (A) Selected MR images. Sagittal T1-weighted (top row) and coronal T2-weighted (bottom row) images of patient 1 (i and i0), patient 2 (ii and ii0), patient 3 (iii and iii0), and patient 4 (iv and iv0) are shown. The images were obtained at age 25 months for patient 1, 16 months for patient 2, 25 months for patient 3, and 41 months for patient 4. All patients have simplified gyral pattern, which is mild in patients 1, 3, and 4, and is moderate in patient 2. The corpus callosum is healthy in patient 1, normally developed but diffusely thinned in patient 2, healthy in patient 3, and mildly hypoplastic in patient 4. Patients 1 and 3 have normally developed anterior commissure (arrows in i, ii0, iii, and iii0), but patients 2 and 4 have an absence of the anterior commissure. Patient 2 has moderate ventriculomegaly. (B) The schematic showing the location of the identified variants in the coding exons and the corresponding parts of the CELF2 protein with three RNA-recognition motifs (RRM1–3) (Ladd and Cooper, 2004). The affected residues are within a known nuclear-localization signal (NLS) in RRM3. (C) Boxplots showing CELFs expression in human cortical NPCs (PAX6+ and SOX2+) (single-cell RNA-seq dataset from Pollen et al., 2015). (D) Temporal quantitative real-time PCR analysis of the expression of CELF1 and CELF2 in the developing mouse cortices at the indicated time points. n = 3 samples at each time point. (E) Confocal images of HEK293 cells expressing EGFP-CELF2 (green) that carry different pathogenic variants and counterstained for Hoechst 33258 (blue). ‘‘N’’ denotes the nucleus. Dashed white lines denote nuclei (blue). The intensity of CELF2 immunosignals across individual cells is shown at the bottom with the nucleus region labeled by the blue bar. Scale bar, 10 mm. (F) RT-PCR analysis of exon-skipping events for PPP1R mRNA and TRAF3 mRNA in HEK293 cells transfected with empty vector (control), wild-type (WT), or mutant CELF2. (G) Quantifications of the percentage of transcripts with the skipped exon. One-way ANOVA with Tukey’s test. n = 3 experiments. (H) The quantitative real-time PCR enrichment analysis of GAPDH mRNA and MCL mRNA by RIP with control IgG or an anti-EGFP antibody in HEK293 cells transfected with EGFP-tagged CELF2-NES or mutant CELF2 (normalized to the total RNA input). n = 3 experiments. Data are presented as means ± SEM. Each symbol represents one experiment.

    Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Sox2 (goat) R&D Systems AF2018 bIII-tubulin (mouse) Biolegend 801201 CELF2 (rabbit) Abcam ab186430 CELF2 (mouse) Santa Cruz Biotechnology sc47731 Ki67 (mouse) BD Biosciences 550609 CSDE1 (rabbit) Abcam ab201688 LaminB1 (mouse) Abcam ab8982 Lap2 (mouse) BD Biosciences 611000 Satb2 (mouse) Abcam ab51502 Tbr2 (rabbit) Abcam ab183991 Tbr2 (rat) Invitrogen 14-4875-80 Pax6 (rabbit) Biolegend 901301 pH3 (mouse) Invitrogen MA5-15220 pH3 (mouse) Cell Signaling 9706s GFP (mouse) Abcam ab290 GFP (chicken) EMD Millipore AB16901 GAPDH (rabbit) Sigma G9545 Dcp1a (mouse) Abnova H00055802-M06 4E-T Abnova H00056478-B01 Bacterial strains pEGFP-C1 Clontech Discontinued pCAGIG Matsuda and Cepko, 2004 Addgene (11159) pSUPER OligoEngine VEC-pBS-0002 Chemicals, peptides, and recombinant proteins Q5 Site-Directed Mutagenesis Kit NEB E0554S Polyethylenimine Sigma Aldrich 765090 FGF2 Corning CB-40060A B27 Thermo Fisher 17504044 Penicillin-streptomycin Wisent 450-201-EL L-glutamine Wisent 609-065-EL DMEM Thermo Fisher 11995065 Neurobasal Thermo Fisher 21103049 FBS Wisent 098-150 Lipofectamine Stem Thermo Fisher STEM00015 Laminin Corning 354232 Poly-D-Lysine Sigma Aldrich P6407 Trizol Invitrogen LS15596026 Maxima H Minus cDNA Synthesis Kit Thermo Fisher M1681 Phusion High Fidelity PCR Master Mix Thermo Fisher F531 PerfeCTa SYBR Green FastMix Quanta Bio 95072 Bovine Serum Albumin (BSA) BioShop 005PJD Bovine Serum Albumin (BSA) Jackson Immunoresearch 005PEF Hoechst 33258 Sigma Aldrich 94403 (Continued on next page) Cell Reports 35, 109226, June 8, 2021 e1

    Techniques: Expressing, RNA Sequencing, Real-time Polymerase Chain Reaction, Labeling, Reverse Transcription Polymerase Chain Reaction, Transfection, Plasmid Preparation, Control, Mutagenesis

    Figure 2. Attenuation of Hippo Signaling Promotes Nuclear Accumulation of YAP in Epiblast (A) Distribution of YAP and SOX2 proteins in early, mid and late blastocyst stage embryos. Nuclear YAP signals gradually increase in the forming epiblast. Dashed lines indicate SOX2-positive ICM or epiblast cells. (A0) Enlargement of (A). (B) Quantification of nuclear YAP signals.

    Journal: Developmental cell

    Article Title: Epiblast Formation by TEAD-YAP-Dependent Expression of Pluripotency Factors and Competitive Elimination of Unspecified Cells.

    doi: 10.1016/j.devcel.2019.05.024

    Figure Lengend Snippet: Figure 2. Attenuation of Hippo Signaling Promotes Nuclear Accumulation of YAP in Epiblast (A) Distribution of YAP and SOX2 proteins in early, mid and late blastocyst stage embryos. Nuclear YAP signals gradually increase in the forming epiblast. Dashed lines indicate SOX2-positive ICM or epiblast cells. (A0) Enlargement of (A). (B) Quantification of nuclear YAP signals.

    Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Goat polyclonal anti-Sox2 antibody R&D Systems Cat# AF2018; RRID: AB_355110 Rabbit monoclonal anti-Tead1 antibody Cell Signaling Technology Cat# D9X2L; RRID: AB_2797873 Rabbit monoclonal anti-Yap1 antibody Cell Signaling Technology Cat# D8H1X; RRID: AB_2650491 Mouse monoclonal anti-Oct3/4 antibody MBL Cat# 2F12; RRID: AB_1953021 Rabbit polyclonal anti-Nanog antibody ReproCell Cat# RCAB001P; RRID: AB_2616320 Rabbit polyclonal anti-p-Yap(S127) antibody Cell Signaling Technology Cat# 4911; RRID: AB_2218913 Rabbit monoclonal anti-active-Yap antibody abcam Cat# ab205270 Rabbit polyclonal anti-Amot antibody Hirate et. al. (2013) Rabbit polyclonal anti-Amotl2 antibody Sugihara-Mizuno et al. (2007) Rabbit polyclonal anti-Cleaved-Caspase3 antibody Cell Signaling Technology Cat# 9611; RRID: AB_330302 Goat polyclonal anti-Sox17 antibody R&D Systems Cat# AF1924; RRID: AB_355060 Rabbit monoclonal anti-Myc antibody Abcam Cat# ab32072; RRID: AB_731658 Mouse monoclonal anti-Cdx2 antibody BioGenex Laboratories Cat# B-MU392AUC Mouse monoclonal anti-Tbp antibody abcam Cat# ab51841; RRID: AB_945758 Mouse monoclonal anti-Esrrb antibody Perseus Proteomics Cat# pp-H6705-00; RRID: AB_2100412 Alexa 488-conjugated donkey anti-goat antibody Thermo Fisher Scientific Cat# A-11055; RRID: AB_142672 Alexa 594-conjugated donkey anti-mouse antibody Thermo Fisher Scientific Cat# A-21203; RRID: AB_141633 Alexa 647-conjugated donkey anti-rabbit antibody Thermo Fisher Scientific Cat# A-31573; RRID: AB_2536183 Chemicals, Peptides, and Recombinant Proteins CHIR99021 ChemScene CS-0181 PD0325901 Cayman 13034 c-Myc inhibitor Calbiochem 475956 Z-VAD-FMK Peptide Institute 3118-v KSOM ARK-Resource I0BAIK200 M2 Sigma-Aldrich M7167 Hyaluronidase Sigma-Aldrich H4272 Opti-MEM I Thermo Fisher Scientific 31985062 PMSG ASKA Animal Health Gonatropin hCG ASKA Animal Health Serotropin Mineral oil Sigma-Aldrich M8410 Hoechst 33342 Dojindo EW189 Alt-R S.p.

    Techniques:

    Figure 3. TEAD Activity Promotes Pluripotency Factor Expression in Forming Epiblast (A–F) Correlations between signals of pluripotency factors and nuclear YAP. Signal quantification of pluripotency factors, SOX2 (A), OCT3/4 (C), and NANOG (E), in SOX2-positive ICM or epiblast cells. Relationships between signals of nuclear YAP and pluripotency factors, SOX2 (B), OCT3/4 (D), and NANOG (F). Data from early, mid, and late blastocyst stages are labeled with yellow, orange, and red colors, respectively. Correlation coefficients and p values are shown above the graphs. (G–I) TEAD activity is required for strong expression of pluripotency factors. (G) Z-VAD-treated WT 5 Tead1–/– embryo showing weaker expression of plurip- otency factors, SOX2 and NANOG, in Tead1–/– cells. Yellow and cyan dashed lines indicate wild-type and Tead1–/– cells, respectively. Scale bar represents 20 mm. Quantification of SOX2 (H) and NANOG (I) signals in WT and Tead1–/– cells at late blastocyst stage. Data are represented as the mean ± SD. One-way ANOVA followed by Dunn’s multiple comparison test (A, C, and E), non-parametric Spearman’s rank correlation (B, D, and F), Mann-Whitney U test (H and I). *p < 0.05, ***p < 0.001, ****p < 0.0001. See also Figure S4.

    Journal: Developmental cell

    Article Title: Epiblast Formation by TEAD-YAP-Dependent Expression of Pluripotency Factors and Competitive Elimination of Unspecified Cells.

    doi: 10.1016/j.devcel.2019.05.024

    Figure Lengend Snippet: Figure 3. TEAD Activity Promotes Pluripotency Factor Expression in Forming Epiblast (A–F) Correlations between signals of pluripotency factors and nuclear YAP. Signal quantification of pluripotency factors, SOX2 (A), OCT3/4 (C), and NANOG (E), in SOX2-positive ICM or epiblast cells. Relationships between signals of nuclear YAP and pluripotency factors, SOX2 (B), OCT3/4 (D), and NANOG (F). Data from early, mid, and late blastocyst stages are labeled with yellow, orange, and red colors, respectively. Correlation coefficients and p values are shown above the graphs. (G–I) TEAD activity is required for strong expression of pluripotency factors. (G) Z-VAD-treated WT 5 Tead1–/– embryo showing weaker expression of plurip- otency factors, SOX2 and NANOG, in Tead1–/– cells. Yellow and cyan dashed lines indicate wild-type and Tead1–/– cells, respectively. Scale bar represents 20 mm. Quantification of SOX2 (H) and NANOG (I) signals in WT and Tead1–/– cells at late blastocyst stage. Data are represented as the mean ± SD. One-way ANOVA followed by Dunn’s multiple comparison test (A, C, and E), non-parametric Spearman’s rank correlation (B, D, and F), Mann-Whitney U test (H and I). *p < 0.05, ***p < 0.001, ****p < 0.0001. See also Figure S4.

    Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Goat polyclonal anti-Sox2 antibody R&D Systems Cat# AF2018; RRID: AB_355110 Rabbit monoclonal anti-Tead1 antibody Cell Signaling Technology Cat# D9X2L; RRID: AB_2797873 Rabbit monoclonal anti-Yap1 antibody Cell Signaling Technology Cat# D8H1X; RRID: AB_2650491 Mouse monoclonal anti-Oct3/4 antibody MBL Cat# 2F12; RRID: AB_1953021 Rabbit polyclonal anti-Nanog antibody ReproCell Cat# RCAB001P; RRID: AB_2616320 Rabbit polyclonal anti-p-Yap(S127) antibody Cell Signaling Technology Cat# 4911; RRID: AB_2218913 Rabbit monoclonal anti-active-Yap antibody abcam Cat# ab205270 Rabbit polyclonal anti-Amot antibody Hirate et. al. (2013) Rabbit polyclonal anti-Amotl2 antibody Sugihara-Mizuno et al. (2007) Rabbit polyclonal anti-Cleaved-Caspase3 antibody Cell Signaling Technology Cat# 9611; RRID: AB_330302 Goat polyclonal anti-Sox17 antibody R&D Systems Cat# AF1924; RRID: AB_355060 Rabbit monoclonal anti-Myc antibody Abcam Cat# ab32072; RRID: AB_731658 Mouse monoclonal anti-Cdx2 antibody BioGenex Laboratories Cat# B-MU392AUC Mouse monoclonal anti-Tbp antibody abcam Cat# ab51841; RRID: AB_945758 Mouse monoclonal anti-Esrrb antibody Perseus Proteomics Cat# pp-H6705-00; RRID: AB_2100412 Alexa 488-conjugated donkey anti-goat antibody Thermo Fisher Scientific Cat# A-11055; RRID: AB_142672 Alexa 594-conjugated donkey anti-mouse antibody Thermo Fisher Scientific Cat# A-21203; RRID: AB_141633 Alexa 647-conjugated donkey anti-rabbit antibody Thermo Fisher Scientific Cat# A-31573; RRID: AB_2536183 Chemicals, Peptides, and Recombinant Proteins CHIR99021 ChemScene CS-0181 PD0325901 Cayman 13034 c-Myc inhibitor Calbiochem 475956 Z-VAD-FMK Peptide Institute 3118-v KSOM ARK-Resource I0BAIK200 M2 Sigma-Aldrich M7167 Hyaluronidase Sigma-Aldrich H4272 Opti-MEM I Thermo Fisher Scientific 31985062 PMSG ASKA Animal Health Gonatropin hCG ASKA Animal Health Serotropin Mineral oil Sigma-Aldrich M8410 Hoechst 33342 Dojindo EW189 Alt-R S.p.

    Techniques: Activity Assay, Expressing, Labeling, Comparison, MANN-WHITNEY

    Figure 4. Endogenous Cell Competition Triggered by TEAD Activity Takes Place at Mid-Blastocyst Stage (A) Wild-type embryos showing cleaved caspase 3 (cl. CASP3) signals in ICM or epiblast. (B) Quantification of apoptotic cells in the ICM or epiblast. (C) Variation in nuclear YAP and SOX2 signals within an embryo. White and yellow arrowheads indicate the cells with strong and weak nuclear YAP or SOX2 signals, respectively. At the late blastocyst stage, the signals are less variable. (D) Quantification of nuclear YAP and SOX2 signals showing strong variation in the signal intensities within individual embryos at mid-blastocyst stage. The data derived from different embryos are labeled with different colors. (E) Cleaved caspase 3-positive cells showing weak nuclear YAP or SOX2 signals (yellow arrowheads). (F and G) Quantification of SOX2 (F) and nuclear YAP (G) signals. (legend continued on next page)

    Journal: Developmental cell

    Article Title: Epiblast Formation by TEAD-YAP-Dependent Expression of Pluripotency Factors and Competitive Elimination of Unspecified Cells.

    doi: 10.1016/j.devcel.2019.05.024

    Figure Lengend Snippet: Figure 4. Endogenous Cell Competition Triggered by TEAD Activity Takes Place at Mid-Blastocyst Stage (A) Wild-type embryos showing cleaved caspase 3 (cl. CASP3) signals in ICM or epiblast. (B) Quantification of apoptotic cells in the ICM or epiblast. (C) Variation in nuclear YAP and SOX2 signals within an embryo. White and yellow arrowheads indicate the cells with strong and weak nuclear YAP or SOX2 signals, respectively. At the late blastocyst stage, the signals are less variable. (D) Quantification of nuclear YAP and SOX2 signals showing strong variation in the signal intensities within individual embryos at mid-blastocyst stage. The data derived from different embryos are labeled with different colors. (E) Cleaved caspase 3-positive cells showing weak nuclear YAP or SOX2 signals (yellow arrowheads). (F and G) Quantification of SOX2 (F) and nuclear YAP (G) signals. (legend continued on next page)

    Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Goat polyclonal anti-Sox2 antibody R&D Systems Cat# AF2018; RRID: AB_355110 Rabbit monoclonal anti-Tead1 antibody Cell Signaling Technology Cat# D9X2L; RRID: AB_2797873 Rabbit monoclonal anti-Yap1 antibody Cell Signaling Technology Cat# D8H1X; RRID: AB_2650491 Mouse monoclonal anti-Oct3/4 antibody MBL Cat# 2F12; RRID: AB_1953021 Rabbit polyclonal anti-Nanog antibody ReproCell Cat# RCAB001P; RRID: AB_2616320 Rabbit polyclonal anti-p-Yap(S127) antibody Cell Signaling Technology Cat# 4911; RRID: AB_2218913 Rabbit monoclonal anti-active-Yap antibody abcam Cat# ab205270 Rabbit polyclonal anti-Amot antibody Hirate et. al. (2013) Rabbit polyclonal anti-Amotl2 antibody Sugihara-Mizuno et al. (2007) Rabbit polyclonal anti-Cleaved-Caspase3 antibody Cell Signaling Technology Cat# 9611; RRID: AB_330302 Goat polyclonal anti-Sox17 antibody R&D Systems Cat# AF1924; RRID: AB_355060 Rabbit monoclonal anti-Myc antibody Abcam Cat# ab32072; RRID: AB_731658 Mouse monoclonal anti-Cdx2 antibody BioGenex Laboratories Cat# B-MU392AUC Mouse monoclonal anti-Tbp antibody abcam Cat# ab51841; RRID: AB_945758 Mouse monoclonal anti-Esrrb antibody Perseus Proteomics Cat# pp-H6705-00; RRID: AB_2100412 Alexa 488-conjugated donkey anti-goat antibody Thermo Fisher Scientific Cat# A-11055; RRID: AB_142672 Alexa 594-conjugated donkey anti-mouse antibody Thermo Fisher Scientific Cat# A-21203; RRID: AB_141633 Alexa 647-conjugated donkey anti-rabbit antibody Thermo Fisher Scientific Cat# A-31573; RRID: AB_2536183 Chemicals, Peptides, and Recombinant Proteins CHIR99021 ChemScene CS-0181 PD0325901 Cayman 13034 c-Myc inhibitor Calbiochem 475956 Z-VAD-FMK Peptide Institute 3118-v KSOM ARK-Resource I0BAIK200 M2 Sigma-Aldrich M7167 Hyaluronidase Sigma-Aldrich H4272 Opti-MEM I Thermo Fisher Scientific 31985062 PMSG ASKA Animal Health Gonatropin hCG ASKA Animal Health Serotropin Mineral oil Sigma-Aldrich M8410 Hoechst 33342 Dojindo EW189 Alt-R S.p.

    Techniques: Activity Assay, Derivative Assay, Labeling

    Figure 5. Pluripotency and MYC Regulate Cell Competition Downstream of TEAD Activity (A) Distribution of wild-type (yellow dashed lines) and Tead1–/– cells (cyan dashed lines) in the epiblast of wild-type 5 Tead1–/– embryos. 2i-treatment suppressed elimination of Tead1–/– cells. (B) Quantification of percentages of Tead1–/– cells in the epiblast. (C) Reduction of cleaved caspase 3-positive cells in 2i-treated wild-type embryos. (D) Distribution of SOX2 and YAP in 2i-treated wild-type embryos. Dashed lines indicate epiblast.

    Journal: Developmental cell

    Article Title: Epiblast Formation by TEAD-YAP-Dependent Expression of Pluripotency Factors and Competitive Elimination of Unspecified Cells.

    doi: 10.1016/j.devcel.2019.05.024

    Figure Lengend Snippet: Figure 5. Pluripotency and MYC Regulate Cell Competition Downstream of TEAD Activity (A) Distribution of wild-type (yellow dashed lines) and Tead1–/– cells (cyan dashed lines) in the epiblast of wild-type 5 Tead1–/– embryos. 2i-treatment suppressed elimination of Tead1–/– cells. (B) Quantification of percentages of Tead1–/– cells in the epiblast. (C) Reduction of cleaved caspase 3-positive cells in 2i-treated wild-type embryos. (D) Distribution of SOX2 and YAP in 2i-treated wild-type embryos. Dashed lines indicate epiblast.

    Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Goat polyclonal anti-Sox2 antibody R&D Systems Cat# AF2018; RRID: AB_355110 Rabbit monoclonal anti-Tead1 antibody Cell Signaling Technology Cat# D9X2L; RRID: AB_2797873 Rabbit monoclonal anti-Yap1 antibody Cell Signaling Technology Cat# D8H1X; RRID: AB_2650491 Mouse monoclonal anti-Oct3/4 antibody MBL Cat# 2F12; RRID: AB_1953021 Rabbit polyclonal anti-Nanog antibody ReproCell Cat# RCAB001P; RRID: AB_2616320 Rabbit polyclonal anti-p-Yap(S127) antibody Cell Signaling Technology Cat# 4911; RRID: AB_2218913 Rabbit monoclonal anti-active-Yap antibody abcam Cat# ab205270 Rabbit polyclonal anti-Amot antibody Hirate et. al. (2013) Rabbit polyclonal anti-Amotl2 antibody Sugihara-Mizuno et al. (2007) Rabbit polyclonal anti-Cleaved-Caspase3 antibody Cell Signaling Technology Cat# 9611; RRID: AB_330302 Goat polyclonal anti-Sox17 antibody R&D Systems Cat# AF1924; RRID: AB_355060 Rabbit monoclonal anti-Myc antibody Abcam Cat# ab32072; RRID: AB_731658 Mouse monoclonal anti-Cdx2 antibody BioGenex Laboratories Cat# B-MU392AUC Mouse monoclonal anti-Tbp antibody abcam Cat# ab51841; RRID: AB_945758 Mouse monoclonal anti-Esrrb antibody Perseus Proteomics Cat# pp-H6705-00; RRID: AB_2100412 Alexa 488-conjugated donkey anti-goat antibody Thermo Fisher Scientific Cat# A-11055; RRID: AB_142672 Alexa 594-conjugated donkey anti-mouse antibody Thermo Fisher Scientific Cat# A-21203; RRID: AB_141633 Alexa 647-conjugated donkey anti-rabbit antibody Thermo Fisher Scientific Cat# A-31573; RRID: AB_2536183 Chemicals, Peptides, and Recombinant Proteins CHIR99021 ChemScene CS-0181 PD0325901 Cayman 13034 c-Myc inhibitor Calbiochem 475956 Z-VAD-FMK Peptide Institute 3118-v KSOM ARK-Resource I0BAIK200 M2 Sigma-Aldrich M7167 Hyaluronidase Sigma-Aldrich H4272 Opti-MEM I Thermo Fisher Scientific 31985062 PMSG ASKA Animal Health Gonatropin hCG ASKA Animal Health Serotropin Mineral oil Sigma-Aldrich M8410 Hoechst 33342 Dojindo EW189 Alt-R S.p.

    Techniques: Activity Assay