Journal: Frontiers in Cell and Developmental Biology

Article Title: ESRRB Facilitates the Conversion of Trophoblast-Like Stem Cells From Induced Pluripotent Stem Cells by Directly Regulating CDX2

doi: 10.3389/fcell.2021.712224

Figure Lengend Snippet: Antibodies used in this experiment.

Article Snippet: CDX2 , Bioss , bs-6694R.

Techniques:

Journal: Cell stem cell

Article Title: TFAP2C- and p63-Dependent Networks Sequentially Rearrange Chromatin Landscapes to Drive Human Epidermal Lineage Commitment

doi: 10.1016/j.stem.2018.12.012

Figure Lengend Snippet: (A) Schematic representation of the piggyBac TetO-TFAP2C inducible expression system in H9 hESC. TRE, tetracycline-responsive element; rtTA, reverse tetracycline transactivator.

Article Snippet: Human TFAP2C cDNA , Sino Biological Inc. , Cat #HG13115-G.

Techniques: Expressing

Journal: Cell stem cell

Article Title: TFAP2C- and p63-Dependent Networks Sequentially Rearrange Chromatin Landscapes to Drive Human Epidermal Lineage Commitment

doi: 10.1016/j.stem.2018.12.012

Figure Lengend Snippet: (A) Schematic diagram shows the experimental procedure to study whether TFAP2C-induced surface ectoderm progenitor cells (TetO-TFAP2C-D7) can further differentiate into functional keratinocytes (TetO-TFAP2C-KC) in maturation medium.

Article Snippet: Human TFAP2C cDNA , Sino Biological Inc. , Cat #HG13115-G.

Techniques: Functional Assay

Journal: Cell stem cell

Article Title: TFAP2C- and p63-Dependent Networks Sequentially Rearrange Chromatin Landscapes to Drive Human Epidermal Lineage Commitment

doi: 10.1016/j.stem.2018.12.012

Figure Lengend Snippet: (A) Schematic illustration of the approach to functionally study p63 via CRISPR/Cas9-mediated gene knockout (KO) during TFAP2C-induced epidermal differentiation.

Article Snippet: Human TFAP2C cDNA , Sino Biological Inc. , Cat #HG13115-G.

Techniques: CRISPR, Gene Knockout

Journal: Cell stem cell

Article Title: TFAP2C- and p63-Dependent Networks Sequentially Rearrange Chromatin Landscapes to Drive Human Epidermal Lineage Commitment

doi: 10.1016/j.stem.2018.12.012

Figure Lengend Snippet: (A) Gene expression changes of TFAP2C and p63 during TFAP2C-induced epidermal differentiation.

Article Snippet: Human TFAP2C cDNA , Sino Biological Inc. , Cat #HG13115-G.

Techniques: Expressing

Journal: Cell stem cell

Article Title: TFAP2C- and p63-Dependent Networks Sequentially Rearrange Chromatin Landscapes to Drive Human Epidermal Lineage Commitment

doi: 10.1016/j.stem.2018.12.012

Figure Lengend Snippet: A proposed model depicts the identified chromatin states and feedback regulation between TFAP2C- and p63-centered TF regulatory networks driving the chromatin transition during epidermal lineage commitment.

Article Snippet: Human TFAP2C cDNA , Sino Biological Inc. , Cat #HG13115-G.

Techniques:

Journal: Cell stem cell

Article Title: TFAP2C- and p63-Dependent Networks Sequentially Rearrange Chromatin Landscapes to Drive Human Epidermal Lineage Commitment

doi: 10.1016/j.stem.2018.12.012

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Human TFAP2C cDNA , Sino Biological Inc. , Cat #HG13115-G.

Techniques: Purification, Recombinant, Knock-Out, Blocking Assay, Plasmid Preparation, Clone Assay, SYBR Green Assay, Magnetic Beads, Staining, Sequencing, CRISPR, Negative Control, shRNA, Expressing, Software

Journal: PloS one

Article Title: Transcription factor TFAP2C regulates major programs required for murine fetal germ cell maintenance and haploinsufficiency predisposes to teratomas in male mice.

doi: 10.1371/journal.pone.0071113

Figure Lengend Snippet: Figure 1. Genes regulated in PGCLCs and ESCs by TFAP2C. Affymetrix microarray gene expression analysis performed with RNA extracted from #1-ctrl; #1-Tfap2c2/2 and #2-ctrl; #2-Tfap2c2/2 ESCs and PGCLCs. (A) Hierarchical clustering. The red bars cluster the ESCs, green bars the Tfap2c2/2PGCLCs and blue bars ctrl PGCLCs, respectively. The shorter the horizontal bar that connects two branches the closer are the populations. (B) Heat map was performed with the probes whose range or variation across all samples was at least 3. Color bar on top codifies the gene expression in log2 scale. Red and blue indicate higher and lower relative expression. (C-D) Pairwise scatter plot of global gene expression in ctrl versus Tfap2c2/2

Article Snippet: Scrambled and TFAP2C siRNA were obtained from Origene (TFAP2C SR304789, Origene Technologies, Rockville, USA).

Techniques: Microarray, Gene Expression, Expressing

Journal: PloS one

Article Title: Transcription factor TFAP2C regulates major programs required for murine fetal germ cell maintenance and haploinsufficiency predisposes to teratomas in male mice.

doi: 10.1371/journal.pone.0071113

Figure Lengend Snippet: Figure 2. Validation of cDNA microarray data. (A) Quantitative RT-PCR of a subset of markers with RNA isolated from ctrl and Tfap2c2/2PGCLCs. Expression levels were normalized to bActin and expression level of ctrl PGCLCs were set to 1. qRT-PCR was performed in biological triplicates. Error bars indicate standard deviation. (B) Quantitative RT-PCR of a subset of markers with RNA isolated from TCam-2 cells after siRNA mediated knockdown of TFAP2C. Expression levels were normalized to GAPDH and expression levels of scrambled-siRNA-transfection were set to 1. qRT-PCR was performed in biological triplicate. Error bars indicate standard deviation. (C) ChIP/qPCR analysis for Tfap2c was performed with four biological replicates of PGCLCs. The qPCR results were calculated with the percentage input method and ChIP analyses with IgG antibody served as control and

Article Snippet: Scrambled and TFAP2C siRNA were obtained from Origene (TFAP2C SR304789, Origene Technologies, Rockville, USA).

Techniques: Biomarker Discovery, Microarray, Quantitative RT-PCR, Isolation, Expressing, Standard Deviation, Knockdown, Transfection, ChIP-qPCR, Control

Journal: PloS one

Article Title: Transcription factor TFAP2C regulates major programs required for murine fetal germ cell maintenance and haploinsufficiency predisposes to teratomas in male mice.

doi: 10.1371/journal.pone.0071113

Figure Lengend Snippet: Figure 3. Teratoma development in Tfap2c heterozygous mice in 129S2/Sv genetic background. (A) Quantitative RT-PCR with RNA isolated from E12.5 genital ridges of wt and Tfap2c+/2 embryos was performed. Expression levels of Tfap2c and p21 were normalized to Gapdh. qRT- PCR was performed in biological duplicates. Error bars indicate standard deviation. (B) Percentage of total tumor incidence in Tfap2c heterozygous 129S2/Sv male mice. The seventh generation in 129S2/Sv shows 82% (n = 51) testicular tumors. Red bar: bilateral cases (35%), blue bar: unilateral tumors (47%). (C) Gross pathology of testicular teratoma in Tfap2c+/2 male mice. (D–G) HE-staining of testicular teratomas of 3–6 month old mice. Tumors show immature glia (D), mature cartilage, muscle (E), respiratory epithelium (F) and squamous epithelium (G). Scale bars: 200 mm. doi:10.1371/journal.pone.0071113.g003

Article Snippet: Scrambled and TFAP2C siRNA were obtained from Origene (TFAP2C SR304789, Origene Technologies, Rockville, USA).

Techniques: Quantitative RT-PCR, Isolation, Expressing, Standard Deviation, Staining

Journal: PloS one

Article Title: Transcription factor TFAP2C regulates major programs required for murine fetal germ cell maintenance and haploinsufficiency predisposes to teratomas in male mice.

doi: 10.1371/journal.pone.0071113

Figure Lengend Snippet: Figure 4. Pluripotency maintaining and differentiation block show EC like nature of lesion. (A–C) IHC staining of E16.5 embryonal testis. (A) SSEA1, (B) OCT3/4 and (C) TFAP2C. Scale bars: 50 mm. (B) Black arrow show cells with large, highly condensated nuclei as well as clearly visible nucleoli. (D) RT-PCR from RNA of microdissected SSEA-1 positive foci of E16.5 testes detecting Nanog, Sox2 and c-Kit. Comparison of microdissected SSEA-1 positive tissue (T) and SSEA-1 negative tissue (wt). Gapdh served as control. doi:10.1371/journal.pone.0071113.g004

Article Snippet: Scrambled and TFAP2C siRNA were obtained from Origene (TFAP2C SR304789, Origene Technologies, Rockville, USA).

Techniques: Blocking Assay, Immunohistochemistry, Reverse Transcription Polymerase Chain Reaction, Comparison, Control

Journal: PloS one

Article Title: Transcription factor TFAP2C regulates major programs required for murine fetal germ cell maintenance and haploinsufficiency predisposes to teratomas in male mice.

doi: 10.1371/journal.pone.0071113

Figure Lengend Snippet: Figure 6. Schematic of the genes and programs regulated by TFAP2C in primordial germ cells. Black arrows indicate pathways transactivated and induced by TFAP2C and black lines with terminal bars indicate pathways repressed by TFAP2C during development of primordial germ cells. Genes listed in respective pathways indicate direct regulation as demonstrated by ChIP analyses. doi:10.1371/journal.pone.0071113.g006

Article Snippet: Scrambled and TFAP2C siRNA were obtained from Origene (TFAP2C SR304789, Origene Technologies, Rockville, USA).

Techniques:

Journal: Life Medicine

Article Title: A CRISPR/Cas9-based kinome screen identifies ErbB signaling as a new regulator of human naïve pluripotency and totipotency

doi: 10.1093/lifemedi/lnad037

Figure Lengend Snippet: Transcriptomic analysis of ErbB family inhibitors induced naïve pluripotency and totipotent features. (A) Heatmap analysis of significantly down-regulated primed pluripotency marker genes (left) and up-regulated naïve pluripotent marker genes (right) upon Afatinib and Erlotinib treatment under the mTeSR condition. (B) Pie chart showing number of up-regulated and down-regulated differentially expressed pan-early embryonic genes under different conditions. PXGL treated, differentially expressed genes shared by PXGL supplemented with Afatinib and that with Erlotinib. (C) Heatmap analysis of totipotency-specific genes under mTeSR, PXGL and PXGL treated with Afatinib and Erlotinib. (D) GSEA enrichment result upon Afatinib treatment under PXGL condition. (E) GSEA enrichment result upon Afatinib treatment under PXGL condition. (F) PPI analysis network showing protein–protein interactions between ErbB family along with their downstream genes and 8CLC hub genes. Interaction score = 0.4. Line thickness indicates the strength of data support. (G) Predicted TF–gene interaction network between ErbB family and 8CLC hub genes. Factor and gene target data derived from the ENCODE ChIP-seq data. Only peak intensity signal < 500 and the predicted regulatory potential score < 1 is used (using BETA minus algorithm). (H) Q-PCR analysis of the expression of ERBB famlily genes and TFAP2C upon TFAP2C transient over-expression under the XF-LCDM condition. N = 2 biological replicates. Error bar indicates SEM. (I) q-PCR analysis of the expression of ERBB family genes and KLF5 upon KLF5 transient over-expression under the XF-LCDM condition. N = 2 biological replicates. Error bar indicates SEM. (J) q-PCR analysis of the expression of ERBB family genes upon SR18662 treatment under the XF-LCDM condition. N = 2 biological replicates. Error bar indicates SEM. (K) q-PCR analysis of the expression of totipotency marker genes upon SR18662 treatment under the XF-LCDM condition. N = 2 biological replicates. Error bar indicates SEM.

Article Snippet: CMV- KLF5 -Neo (Origene, RC202438) and CMV- TFAP2C -Neo (Origene, RC208665) were transduced into hEPSCs via nucleofection.

Techniques: Marker, Protein-Protein interactions, Derivative Assay, ChIP-sequencing, Expressing, Over Expression

Journal: Molecular Oncology

Article Title: Interaction of transcription factor AP‐2 gamma with proto‐oncogene PELP1 promotes tumorigenesis by enhancing RET signaling

doi: 10.1002/1878-0261.12871

Figure Lengend Snippet: PELP1 interacts with TFAP2C. (A, B) Total lysates from MCF7‐control and MCF7‐GFP‐PELP1 cells (A) or ZR75‐control and ZR75‐GFP‐PELP1 cells (B) were subjected to IP using the GFP‐TRAP beads, and the TFAP2C interaction was verified by western blotting ( n = 2). (C) Total lysates from MCF7 and ZR75 cells were subjected to IP using the control IgG or TFAP2C antibody, and the PELP1 interaction was verified by western blotting ( n = 2). (D) Total lysates from MCF7 cells were subjected to GST pull‐down assays using the purified control GST or GST‐PELP1 full‐length proteins expressed in Escherichia coli and the TFAP2C interaction was verified by western blotting. (E) Escherichia coli expressed and purified GST‐PELP1 deletions were used to identify TFAP2C interacting regions in the PELP1 protein using MCF7 total lysates. (F) Schematic representation of GST‐PELP1 deletions utilized in the pull‐down assays.

Article Snippet: MCF7, ZR75, and HEK293T model cell lines were seeded into 24‐well plates and after overnight incubation, transiently transfected with 250 ng AP2‐LUC reporter vector along with PELP1 shRNA vector (SuperArray Bioscience Corporation, Frederick, MD, USA), TFAP2C (GFP‐tagged) expressing vector (RG208665; OriGene, Rockville, MD, USA) or control vector using Turbofect Transfection Reagent (Thermo Fisher Scientific).

Techniques: Western Blot, Purification

Journal: Molecular Oncology

Article Title: Interaction of transcription factor AP‐2 gamma with proto‐oncogene PELP1 promotes tumorigenesis by enhancing RET signaling

doi: 10.1002/1878-0261.12871

Figure Lengend Snippet: PELP1 serves as coactivator of TFAP2C. (A) Top TFs enriched in PELP1‐regulated genes identified using published PELP1 RNA‐seq data. (B) GSEA of target genes of NF‐κB and TFAP2C with PELP1 RNA‐seq data. (C) HEK293T cells were transfected with AP‐2 Luc plasmid along with increasing concentrations of TFAP2C, and 48h after transfection, the reporter activity was measured. (D) HEK293T cells stably expressing either control or PELP1 shRNA were cotransfected with AP‐2 Luc plasmid along with TFAP2C (100 ng), and 48 h after transfection, the reporter activity was measured. PELP1 knockdown was confirmed using western blotting. (E) MCF7 control and MCF7‐GFP‐PELP1 cells were transfected with AP‐2 Luc plasmid, and the reporter activity was measured after 48 h. (F) MCF7 control and MCF7‐PELP1 shRNA cells or ZR75‐control and ZR75‐PELP1 shRNA cells were transfected with AP‐2 Luc plasmid, and the reporter activity was measured after 48 h. (G) GSEA testing correlation of target genes of PELP1 with genes upregulated by TFAP2C and repressed by TFAP2C. (H) Heat map showing changes in expression levels of several TFAP2C‐upregulated and TFAP2C‐repressed genes in PELP1 RNA‐seq data. (I) TFAP2C knockdown was confirmed using western blotting. (J) Total RNA was isolated from MCF7 model cells expressing either PELP1 shRNA or TFAP2C shRNA, and the status of TFAP2C‐upregulated genes was validated using RT–qPCR. Data are shown as the means ± SEM of three experiments. * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001 by Student's t ‐test in C–F and J.

Article Snippet: MCF7, ZR75, and HEK293T model cell lines were seeded into 24‐well plates and after overnight incubation, transiently transfected with 250 ng AP2‐LUC reporter vector along with PELP1 shRNA vector (SuperArray Bioscience Corporation, Frederick, MD, USA), TFAP2C (GFP‐tagged) expressing vector (RG208665; OriGene, Rockville, MD, USA) or control vector using Turbofect Transfection Reagent (Thermo Fisher Scientific).

Techniques: RNA Sequencing Assay, Transfection, Plasmid Preparation, Activity Assay, Stable Transfection, Expressing, shRNA, Western Blot, Isolation, Quantitative RT-PCR

Journal: Molecular Oncology

Article Title: Interaction of transcription factor AP‐2 gamma with proto‐oncogene PELP1 promotes tumorigenesis by enhancing RET signaling

doi: 10.1002/1878-0261.12871

Figure Lengend Snippet: PELP1 is essential for TFAP2C‐mediated oncogenic functions and therapy resistance. (A) Total lysates from MCF7 or ZR75 cells expressing TFAP2C or PELP1 shRNA were analyzed by western blotting for the status of TFAP2C, ERα, and PELP1. Data are shown from two independent experiments. (B) Equal number of MCF7 or ZR75 model cells expressing TFAP2C or/and PELP1 shRNA was plated in six‐well plates and cultured for 14 days, and the number of colonies for each group was counted ( n = 3). (C) Anchorage‐independent growth potential of the MCF7 cells expressing TFAP2C or/and PELP1 shRNA was measured by soft agar colony formation assay. Representative photographs and quantitation of the soft agar colony formation are shown, scale bar = 100 µm. (D) MCF7 or ZR75 model cells expressing TFAP2C or/and PELP1 shRNA were treated with E2 + Fulvestrant (ICI) for 5 days and then cultured for seven subsequent days. The number of colonies for each group was counted. Data are shown as the means ± SEM of three experiments. * P < 0.05; *** P < 0.001; **** P < 0.0001 by Student's t ‐test in B–D.

Article Snippet: MCF7, ZR75, and HEK293T model cell lines were seeded into 24‐well plates and after overnight incubation, transiently transfected with 250 ng AP2‐LUC reporter vector along with PELP1 shRNA vector (SuperArray Bioscience Corporation, Frederick, MD, USA), TFAP2C (GFP‐tagged) expressing vector (RG208665; OriGene, Rockville, MD, USA) or control vector using Turbofect Transfection Reagent (Thermo Fisher Scientific).

Techniques: Expressing, shRNA, Western Blot, Cell Culture, Soft Agar Assay, Quantitation Assay

Journal: Molecular Oncology

Article Title: Interaction of transcription factor AP‐2 gamma with proto‐oncogene PELP1 promotes tumorigenesis by enhancing RET signaling

doi: 10.1002/1878-0261.12871

Figure Lengend Snippet: PELP1 is needed to facilitate optimal histone methyl modifications at the RET promoter by TFAP2C. (A) Schematic representation of TFAP2C‐binding sites in the RET promoter based on published TFAP2C ChIP‐seq data in the RET locus of MCF7 cells. (B) Primary ChIP was conducted using TFAP2C antibody followed by Re‐ChIP using PELP1 antibody. The status of TFAP2C and PELP1 at the RET promoter was determined by qPCR using primers spanning four TFAP2C‐binding sites. (C) Status of the TFAP2C, PELP1, and histone methyl marks H3K4me3 and H3K9me3 in the RET promoter was determined by qPCR using primers spanning the four TFAP2C‐binding sites. Isotype‐matched IgG ChIP was used as control. Data are shown as the means ± SEM of two experiments. * P < 0.05; ** P < 0.01; *** P < 0.001; ****P < 0.0001 by Student's t ‐test in B–C.

Article Snippet: MCF7, ZR75, and HEK293T model cell lines were seeded into 24‐well plates and after overnight incubation, transiently transfected with 250 ng AP2‐LUC reporter vector along with PELP1 shRNA vector (SuperArray Bioscience Corporation, Frederick, MD, USA), TFAP2C (GFP‐tagged) expressing vector (RG208665; OriGene, Rockville, MD, USA) or control vector using Turbofect Transfection Reagent (Thermo Fisher Scientific).

Techniques: Binding Assay, ChIP-sequencing

Journal: Molecular Oncology

Article Title: Interaction of transcription factor AP‐2 gamma with proto‐oncogene PELP1 promotes tumorigenesis by enhancing RET signaling

doi: 10.1002/1878-0261.12871

Figure Lengend Snippet: PELP1 is needed for optimal activation of RET signaling by TFAP2C. (A) Total RNA was isolated from MCF7 model cells expressing either PELP1 shRNA or TFAP2C shRNA, and the expression of RET and GFRA1 genes was validated using RT–qPCR. (B) Total lysates from MCF7 cells expressing TFAP2C shRNA or PELP1 shRNA were analyzed by western blotting for the status of RET and GFRA1. (C) Total lysates from MCF7 or ZR75 model cells expressing TFAP2C or/and PELP1 shRNA were analyzed by western blotting for the status of RET and GFRA1. (D) Total lysates from MCF7 or ZR75 model cells expressing TFAP2C or/and PELP1 shRNA were stimulated with E2 for 24h, and the status of RET was analyzed by western blotting. (E) Total lysates from MCF7 or ZR75 model cells expressing TFAP2C or/and PELP1 shRNA were analyzed by western blotting for the status of AKT and MAPK pathways. (F) Total lysates from MCF7 model cells expressing RET‐shRNA were analyzed by western blotting for the status of phospho‐AKT and phospho‐ ERK1/2. (G) Effect of RET inhibitor BLU667 (5 μ m ) on AKT and ERK1/2 signaling using MCF7 or ZR75 model cells expressing control vector or TFAP2C was measured by western blotting. (H) MCF7 model cells expressing vector or TFAP2C or PELP1 shRNA were plated in 96‐well plates, and cell viability was measured using an MTT assay in the presence or absence of the RET inhibitor BLU667. Data are shown as the means ± SEM of three experiments. * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001 by Student's t ‐test in A and H.

Article Snippet: MCF7, ZR75, and HEK293T model cell lines were seeded into 24‐well plates and after overnight incubation, transiently transfected with 250 ng AP2‐LUC reporter vector along with PELP1 shRNA vector (SuperArray Bioscience Corporation, Frederick, MD, USA), TFAP2C (GFP‐tagged) expressing vector (RG208665; OriGene, Rockville, MD, USA) or control vector using Turbofect Transfection Reagent (Thermo Fisher Scientific).

Techniques: Activation Assay, Isolation, Expressing, shRNA, Quantitative RT-PCR, Western Blot, Plasmid Preparation, MTT Assay

Journal: Molecular Oncology

Article Title: Interaction of transcription factor AP‐2 gamma with proto‐oncogene PELP1 promotes tumorigenesis by enhancing RET signaling

doi: 10.1002/1878-0261.12871

Figure Lengend Snippet: PELP1 is needed for TFAP2C‐mediated BC progression. SCID mice implanted with an E2 pellet were injected with ZR75‐control, ZR75‐TFAP2C, ZR75‐PELP1 shRNA, ZR75‐TFAP2C‐PELP1 shRNA cells into the mammary fat pad, and tumor growth was measured at 5 days intervals. (A) Tumor volume ( n = 6) is shown in the graph. * P < 0.05; **** P < 0.0001 by two‐way ANOVA with Tukey for post hoc test. B,C, Status of Ki‐67 expression as a marker of proliferation and RET was analyzed using IHC. Quantification of Ki‐67 and RET expression is shown (B). (C) Representative IHC images of Ki‐67 and RET are shown. Scale bar = 100µm. Data are shown as the means ± SEM. *** P < 0.001; **** P < 0.0001 by Student's t ‐test.

Article Snippet: MCF7, ZR75, and HEK293T model cell lines were seeded into 24‐well plates and after overnight incubation, transiently transfected with 250 ng AP2‐LUC reporter vector along with PELP1 shRNA vector (SuperArray Bioscience Corporation, Frederick, MD, USA), TFAP2C (GFP‐tagged) expressing vector (RG208665; OriGene, Rockville, MD, USA) or control vector using Turbofect Transfection Reagent (Thermo Fisher Scientific).

Techniques: Injection, shRNA, Expressing, Marker