embryonic kidney cell line hek293t  (ATCC)

Journal: Cell

Article Title: Progestogen-driven B7-H4 contributes to onco-fetal immune tolerance.

doi: 10.1016/j.cell.2024.06.012

Figure Lengend Snippet: Figure 5. PR signaling promotes B7-H4 transcription via binding to the 58 kb enhancer (A) mRNA level of B7-H4 in T-47D cells treated with 100 nM progesterone for 24 h. (B and C) PR ChIP-seq signal tracks and enrichment scores at the indicated region in T-47D cells. (D) H3K27ac, H3K4me1, and H3K4me3 ChIP-seq and ATAC-seq signal tracks in T-47D cells. (E) Hi-C and H3K27ac ChIP-seq signal tracks in T-47D cells treated with R5020. (F) 3C-qPCR for the promoter and putative enhancers in T-47D cells with or without progesterone treatment. P3 is the anchor primer. (G) Luciferase-reporter assay in HEK293T showing increased transcription by adding the putative enhancer to the promoter region. (H) B7-H4 expression after progesterone treatment (100 nM) in T-47D clones with or without 58 kb enhancer region. (I) H3K27ac and PR ChIP-seq signal tracks in ZR-75-1 cells. (J) H3K27ac ChIP-seq signal tracks in human cancer tissues. (K) H3K27ac and H3K4me1 ChIP-seq and ATAC-seq signal tracks in human placental tissues.

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Human first trimester placenta villi scRNA-seq dataset Shannon et al.57 GSE174481 Bulk RNA-seq human breast cancer treated with RU486 Elia et al.68 GSE212690 PR ChIP-seq in T-47D Mohammed et al.115 GSE68359 ATAC-seq and H3K ChIP-seq in T-47D Chan et al.116 GSE107176 Hi-C of T-47D Le Dily et al.117 GSE53463 H3K27ac ChIP-Seq in ZR-75-1 Bi et al.118 GSE128460 PR ChIP-Seq in ZR-75-1 Need et al.119 PRJNA252531 H3K27ac ChIP-Seq in human tumor tissues Singh et al.120 GSE114737 H3K27ac ChIP-Seq in human tumor tissues Reddy et al.121 GSE152885 H3K27ac and H3K4me1 ChIP-Seq in Human placenta Schultz et al.122 GSE18927 PR ChIP-Seq in human endometrium La Greca et al.123 GSE132713 H3K27ac ChIP-seq in T-47D Takaku et al.124 GSE130703 BRD4 ChIP-Seq in T47D Shu et al.125 GSE63584 Bulk RNA-seq of trophoblast cells Kallol et al.126 GSE216484 Bulk RNA-seq of trophoblast cells Liu et al.127 GSE150616 Single nuclei RNA-seq of mouse placental labyrinth Marsh et al.61 GSE152248 Experimental Models: Cell Lines HEK293T ATCC CRL-3126

Techniques: Binding Assay, ChIP-sequencing, Hi-C, Luciferase, Reporter Assay, Expressing, Clone Assay

Journal: BMC Cancer

Article Title: FLI1 induces erythroleukemia through opposing effects on UBASH3A and UBASH3B expression

doi: 10.1186/s12885-024-12075-2

Figure Lengend Snippet: FLI1 binds to the UBASH3B promoter and activates its expression. A The genomic structure of the UBASH3B promoter its indicated derivatives UBASH3B P1, UBASH3B P2, and UBASH3B P1-mut, which were subcloned upstream from the PGL3 luciferase reporter plasmid. B The UBASH3B promoter sequence and its potential FLI1 binding site. C Luciferase activity in HEK293T cells transfected with the UBASH3B P1/P2 and UBASH3B P1-mut luciferase vectors transfected with either FLI1 expression vector MigR1- Fli1 or control plasmid MigR1. D Chromatin immunoprecipitation (ChIp) analysis of the human UBASH3B promoter in HEL erythroleukemic cells for binding to FLI1 by RT-qPCR (top panel). The lower panel shows the gel image for the immunoprecipitated PCR-amplified band relative to the input. P < 0.0001 (****). The full-length gel for Fig. 2D are presented in Supplementary Fig.

Article Snippet: The human leukemia (HEL 92.1.7, K562) and epithelial-like HEK293T (CRL-3216) cell lines were obtained from ATCC (US) and tested negative for mycoplasma.

Techniques: Expressing, Luciferase, Plasmid Preparation, Sequencing, Binding Assay, Activity Assay, Transfection, Control, Chromatin Immunoprecipitation, Quantitative RT-PCR, Immunoprecipitation, Amplification

Journal: BMC Cancer

Article Title: FLI1 induces erythroleukemia through opposing effects on UBASH3A and UBASH3B expression

doi: 10.1186/s12885-024-12075-2

Figure Lengend Snippet: FLI1 interacts with the UBASH3A promoter and reduces its expression. A The genomic structure of the human UBASH3A promoter and its sub-derivatives UBASH3A P1 and UBASH3A P2 as well as their derivative mutant DNA subcloned downstream of the pGL3-basic luciferase reporter plasmid. B The sequence of the UBASH3A promoter and its potential FLI1 binding site. C HEK293T cells were co-transfected with the UBASH3A P1/P2 and mutant (UBASH3A P2-mut) luciferase vectors and either MigR1- Fli1 or control plasmid MigR1. Luciferase activity was determined, as described in materials and methods. D ENCODE data showing the binding of GATA2 to the UBASH3A promoter region—Maximum fold change: 33.3354

Article Snippet: The human leukemia (HEL 92.1.7, K562) and epithelial-like HEK293T (CRL-3216) cell lines were obtained from ATCC (US) and tested negative for mycoplasma.

Techniques: Expressing, Mutagenesis, Luciferase, Plasmid Preparation, Sequencing, Binding Assay, Transfection, Control, Activity Assay