Journal: bioRxiv

Article Title: O-GlcNAc regulates MTA1 transcriptional activity during breast cancer cells genotoxic adaptation

doi: 10.1101/2021.02.08.430201

Figure Lengend Snippet: (a) Breast cancer cells with intrinsic and acquired Adm adaptation contain higher levels of MTA1 and OGT compared with Adm sensitive cells. The expression of indicated proteins in breast cancer cells were analysed by immunoblotting. (b) The PPIs between MTA1, OGT and NuRD components are increased in MDA-MB-231 and MCF-7/ADR cells compared with MCF-7 cells. MTA1 immunoprecipitation was performed and the immunoprecipitated fractions were analysed by immunoblotting by indicated antibodies. (c) The inhibition of O-GlcNAc modification increases the sensitivity of breast cancer cells to Adm and restrains the interaction of MTA1 with OGT and NuRD complex. MCF-7/ADR cells were transfected with OGT siRNA (siOGT) or treated with 100 μM L01 and then incubated with indicated doses of Adm for 48 h. The protein levels were examined by immunoblotting. Vehicle, cells treated with DMSO. The cell viability was assessed through an CCK8 assay. (d) The activation of O-GlcNAc modification increases the adaptation of MCF-7 cells to Adm and elevates the interaction of MTA1 with OGT and NuRD complex. MCF-7 cells were transfected with OGT expression plasmid (pcDNA-OGT) or treated with 100 μM PugNAc and then incubated with indicated doses of Adm for 48 h. The protein levels were examined by immunoblotting. Vehicle, cells treated with DMSO. The cell viability was assessed through an CCK8 assay.

Article Snippet: OGT siRNA (#sc-40781), c-Fos siRNA (#sc-29221) and MTA1 shRNA (#sc-35981-SH) were purchased from Santa Cruz Biotechnology.

Techniques: Expressing, Western Blot, Immunoprecipitation, Inhibition, Modification, Transfection, Incubation, CCK-8 Assay, Activation Assay, Plasmid Preparation

Journal: bioRxiv

Article Title: O-GlcNAc regulates MTA1 transcriptional activity during breast cancer cells genotoxic adaptation

doi: 10.1101/2021.02.08.430201

Figure Lengend Snippet: (a) Left images: Integrative Genomics Viewer (IGV) tracks showing ChIP-seq signal at the promoter regions of c-Fos . Right images: Validation of O-GlcNAc MTA1 binding peaks by ChIP-qPCR. Each bar represents the fold enrichment of binding relative to the input. IgG and random primers that could not specifically bind the indicated gene promoter regions (off target) were used as negative controls. Mock, MCF-7 cells transfected with empty vector. (b) MCF-7/ADR cells were transfected with a reporter vector consisting of luciferase cDNA fused to the c-Fos promoter. The pGL3-basic vector (Vehicle) was used as a control. (c) Effect of MTA1 O-GlcNAc modification on c-Fos transcription and protein levels in MCF-7/ADR cells. The gene mRNA levels were analyzed by qPCR. The protein levels were analyzed by immunoblotting. Vehicle, cells transfected with empty vector. (d) MCF-7/ADR cells were transfected with c-Fos siRNA and then incubated with indicated doses of Adm for 48 h. Vehicle, cells treated with DMSO. The cell viability was assessed through an CCK8 assay. The protein levels were examined by immunoblotting. (e) Proposed model for the MTA1 transcriptional activity regulated by O-GlcNAc modification during breast cancer cells genotoxic adaptation. O-GlcNAc modification enhances the interaction of MTA1 with NuRD complex and chromatin. O-GlcNAc MTA1 preferentially modulates target genes transcription and drives breast cancer cells adaptation during genotoxic stimulation.

Article Snippet: OGT siRNA (#sc-40781), c-Fos siRNA (#sc-29221) and MTA1 shRNA (#sc-35981-SH) were purchased from Santa Cruz Biotechnology.

Techniques: ChIP-sequencing, Biomarker Discovery, Binding Assay, ChIP-qPCR, Transfection, Plasmid Preparation, Luciferase, Control, Modification, Western Blot, Incubation, CCK-8 Assay, Activity Assay

Journal: iScience

Article Title: Growth differentiation factor 15 neutralization does not impact anorexia or survival in lipopolysaccharide-induced inflammation

doi: 10.1016/j.isci.2021.102554

Figure Lengend Snippet:

Article Snippet: Rabbit anti-c-Fos IgG (FD Neurotechnologies) , Santa Cruz Biotechnology , Cat. # sc-52.

Techniques: Control, Recombinant, Enzyme-linked Immunosorbent Assay, Knock-Out, Software