anti zeb1  (Cell Signaling Technology Inc)

Journal: Cell reports

Article Title: Breast cancer cell mesenchymal transition and metastasis directed by DAP5/eIF3d-mediated selective mRNA translation

doi: 10.1016/j.celrep.2023.112646

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Rabbit polyclonal anti-human/mouse Zeb1 antibody , BosterBio , Cat# A00548-1/RRID N/A.

Techniques: Recombinant, Staining, Protease Inhibitor, Virus, Reverse Transcription, SYBR Green Assay, Bicinchoninic Acid Protein Assay, MTT Assay, Software, Imaging

Journal: bioRxiv

Article Title: DNA damage-induced PARP/ALC1 activation leads to Epithelial-to-Mesenchymal transition stimulating homologous recombination

doi: 10.1101/2024.01.16.575847

Figure Lengend Snippet: A . Western blot analyses for ZEB1 after introduction of an shZEB1 expressing vector into different 53BP1-KO clones. B . Relative mRNA level of expression of epithelial and EMT markers in 53BP1-KO clones expressing shZEB1 as compared to the parental cell line (shCTRL). p values from paired comparisons are indicated. NS: non-significant. C . Western blot analyses showing the evolution of the DNA damage marker γH2AX in HCT116 53BP1-KO clones expressing shZEB1 as compared to the control (shCTRL). Also shown are the effects of this expression on ZEB1 and Slug. D . Western blot analyses showing the impact on EMT markers as well as the DNA damage marker γH2AX of forced expression of TWIST1 in PNT1A cells. E . DNA damage recovery assay in HCT116 cells, WT and KO for 53BP1, and depleted or not for ZEB1. Cells were treated (or not, U) with etoposide (2 µM for 24h) before medium change (0 hr) and let to recover for 6 hours. Shown here are western blot analyses for DNA damage markers γ-ATP and γH2AX. F . Western blot analyses for ZEB1, Slug and RAD51 after introduction of a vector expressing either shZEB1 or shSlug into WT or 53BP1-KO HCT116 cells. G . Quantification of γH2AX and RAD51 foci detected by immunofluorescence in WT or 53BP1-KO HCT116 cells expressing either shZEB1 or shSlug. p values from paired comparisons against the control (shCTRL) are indicated. H . Results from Cut&Run experiments using HCT116 cells and either anti-ZEB1 (bottom left) or anti-Slug (bottom right) antibodies for chromatin immunoprecipitation followed by enrichment analyses of two amplified DNA fragments (P1, P2) within the promoter region (ENSEMBL coordinates are indicated) of the RAD51 gene, shown here to span two small regions right upstream of the transcription initiation site (indicated by an arrow). HCT116 cells were either untreated (UT) or transduced with vectors expressing either shZEB1 (bottom left) or shSlug (bottom right). Positive control included an antibody against the histone mark H3K27me3. All PCR signals were normalized to the result obtained with positive antibody control (H3K27m3) and then to an amplified Alu fragment. Significant differences between WT and shZEB1 and shSLUG from multiple t-tests (adjusted) are indicated. * , p < 0.05; ** , p < 0.01; *** , p < 0.001; **** , p < 0.0001.

Article Snippet: ZEB1 (Origene, # TA802298) and Slug (Origene, # OTI1A6) antibodies were used.

Techniques: Western Blot, Expressing, Plasmid Preparation, Clone Assay, Marker, Control, Immunofluorescence, Chromatin Immunoprecipitation, Amplification, Transduction, Positive Control

Journal: bioRxiv

Article Title: DNA damage-induced PARP/ALC1 activation leads to Epithelial-to-Mesenchymal transition stimulating homologous recombination

doi: 10.1101/2024.01.16.575847

Figure Lengend Snippet: A . U2OS cells expressing a GFP-ZEB1 fusion protein were micro-irradiated with laser under the microscope across nuclei and the recruitment of the fluorescent protein was followed and quantified at different time points. Cells were treated or not with Olaparib, a PARP inhibitor, at 5µM. B . Similar experiments were carried out with U2OS cells that had been inactivated for ALC1. C-D . Relative mRNA level of expression of epithelial and EMT markers in 53BP1-KO HEK-Early cells ( C ) and in 53BP1-KO PNT1A cells ( D ) treated with Olaparib at the dose and time indicated, as compared to untreated cells. p values from paired comparisons are indicated. NS: non-significant. E . Western blot analyses of EMT-TFs, Vimentin, RAD51 and γH2AX in 53BP1-KO PNT1A cells before and after treatment with Olaparib. F . Relative mRNA level of expression of epithelial and EMT markers in 53BP1-KO PNT1A cells that were also inactivated for ALC1. p values from paired comparisons are indicated. NS: non-significant.

Article Snippet: ZEB1 (Origene, # TA802298) and Slug (Origene, # OTI1A6) antibodies were used.

Techniques: Expressing, Irradiation, Microscopy, Western Blot