Journal: Cell & Bioscience

Article Title: KDM6B inhibition enhances chemotherapeutic response in small cell lung cancer via epigenetic regulation of apoptosis and ferroptosis

doi: 10.1186/s13578-025-01496-6

Figure Lengend Snippet: Gskj1 enhances H3K27me3 enrichment at EGR3 and suppresses EGR3/c-FOS signaling to promote apoptosis and ferroptosis. A Volcano plot of differentially expressed genes (DEGs) in H69-AR cells treated with Gskj1 versus DMSO, based on RNA-seq analysis (|log₂FC|> 1, P < 0.05). B KEGG pathway enrichment analysis of DEGs showing involvement in apoptosis and tumor progression. C GSEA analysis indicating significant enrichment of pathways related to apoptotic regulation. D Genome-wide H3K27me3 peak signal profiles from ChIP-seq ( GSE150600 ), showing increased H3K27me3 occupancy at promoter regions and ± 3 kb around transcription start sites following KDM6A/B inhibition. E Heatmap of H3K27me3 signal intensity changes across gene bodies and promoters in the presence or absence of KDM6A/B inhibition. F Venn diagram identifying 13 overlapping genes between downregulated RNA-seq DEGs and genes with increased H3K27me3 peaks, including EGR3 and RBFOX3. G Protein–protein interaction network (STRING database) among overlapping candidates and tumor-related genes, showing co-expression of EGR3 and RBFOX3 with c-FOS (confidence > 0.7), as well as predicted co-expression between KDM6B and c-FOS. H H3K27me3 signal heatmaps for candidate genes (EGR3, RBFOX3, SOX2, MYC, TP53, GATA3) showing increased occupancy, especially at the EGR3 promoter and gene body regions following KDM6A/B inhibition. I Peak signal intensity plots further confirming enhanced H3K27me3 enrichment at EGR3 loci. J Western blot validation of EGR3 and c-FOS downregulation following Gskj1 treatment, accompanied by increased H3K27me3 levels. K Co-immunoprecipitation (Co-IP) analysis demonstrating protein–protein interaction between EGR3 and c-FOS. L Dual-luciferase reporter assay showing EGR3 activates the c-FOS promoter, while mutation abolishes this effect

Article Snippet: For each immunoprecipitation reaction, 500–1000 μg of total protein was incubated overnight at 4 °C with 2–5 μg of anti-EGR3 antibody (Abmart, #PK44695) or anti-c-FOS antibody (Abmart, # T56596 ).

Techniques: RNA Sequencing, Genome Wide, ChIP-sequencing, Inhibition, Expressing, Western Blot, Biomarker Discovery, Immunoprecipitation, Co-Immunoprecipitation Assay, Luciferase, Reporter Assay, Mutagenesis

Journal: bioRxiv

Article Title: Dysregulation of neuronal activity-dependent immediate early genes in a mouse model of Angelman syndrome

doi: 10.1101/2025.04.15.648865

Figure Lengend Snippet: Both WT and AS mice were sacrificed, and their age of P9 (before eye opening) and P25 (after eye-opening), visual cortex were carefully removed and processed for immunoblot analysis using antibodies against Arc, Egr1, Egr3, Ube3a, and β-actin. Band intensities of the blots were quantified, normalized against β-actin, and expressed as fold change. A) Immunoblot data. B) Decreased levels of Arc, Egr1, and Egr3 in the AS mice visual cortex at P25 compared to WT mice. Values represented are mean ± SEM with three mice samples in each group. * P < 0.001 compared to P25 WT group (One-way ANOVA followed by post hoc test). Each lane in the immunoblot represents a sample from different mice. C, D) Protein and mRNA levels of Ube3a were significantly increased in the visual cortex of WT mice after eye opening at P25 when compared to before eye opening at P9. C) Quantitation of immunoblot data of Ube3a shown in A. D) Quantitative RT-PCR analysis of Ube3a. In C and D, values are mean ± SEM with 4-6 animals in each group. * P < 0.01 compared to P9 age group ( t -test).

Article Snippet: Mouse monoclonal antibodies against Egr1 (sc-515830), Ube3a (sc-16689), Arc (sc-17839), and Egr3 (sc-390936) were purchased from Santa Cruz Biotechnology.

Techniques: Western Blot, Quantitation Assay, Quantitative RT-PCR

Journal: bioRxiv

Article Title: Dysregulation of neuronal activity-dependent immediate early genes in a mouse model of Angelman syndrome

doi: 10.1101/2025.04.15.648865

Figure Lengend Snippet: Brain sections with visual cortical area obtained from WT and AS mice (P25) were placed on the same slide and subjected to immunofluorescence staining with Arc, Egr1, Egr3, and Ube3a antibody followed by fluorescence-conjugated secondary antibody. Representative images of Ube3a, Arc, Egr1 and Egr3 are shown. Three mice from each group were analysed for immunofluorescence staining. Scale bar: 50 µm.

Article Snippet: Mouse monoclonal antibodies against Egr1 (sc-515830), Ube3a (sc-16689), Arc (sc-17839), and Egr3 (sc-390936) were purchased from Santa Cruz Biotechnology.

Techniques: Immunofluorescence, Staining, Fluorescence

Journal: bioRxiv

Article Title: Dysregulation of neuronal activity-dependent immediate early genes in a mouse model of Angelman syndrome

doi: 10.1101/2025.04.15.648865

Figure Lengend Snippet: Both WT and AS mice were nurtured either in the presence of normal rearing (NR, 12 h light/12 h dark cycle) or dark rearing (DR, constant darkness up to P25), sacrificed at P25, visual cortex were collected and subjected to immunoblot analysis using antibodies against Ube3a, Arc, Egr1, Egr3, and β-actin. Band intensities were quantified, normalized with β-actin, and plotted as fold change. A) Immunoblot data. B) Quantitation data of Arc, Egr1 and Egr3. C) Quantitation data of Ube3a. Values are mean ± SEM with samples from 3 animals in each group. In B, * P < 0.001 compared to the normal rearing NR-WT group (One-way ANOVA followed by post hoc test). Please note that the levels of Egr1, Egr3, and Arc are not significantly different between NR-AS and DR-AS groups. In C, * P <0.001 compared to the NR-WT group ( t -test). Each lane in the immunoblot denotes a sample from different mice.

Article Snippet: Mouse monoclonal antibodies against Egr1 (sc-515830), Ube3a (sc-16689), Arc (sc-17839), and Egr3 (sc-390936) were purchased from Santa Cruz Biotechnology.

Techniques: Western Blot, Quantitation Assay

Journal: bioRxiv

Article Title: Dysregulation of neuronal activity-dependent immediate early genes in a mouse model of Angelman syndrome

doi: 10.1101/2025.04.15.648865

Figure Lengend Snippet: AS mice, along with WT controls, were sacrificed at their age of P9 and P25, and hippocampal tissue was collected and subjected to immunoblot analysis using antibodies against Ube3a, Arc, Egr3, and β-actin. Band intensities of each marker were quantified, normalised, and expressed as fold change. A) Immunoblot data. B) Quantitation of band intensities of Ube3a Arc and Egr3. Arc and Egr3 data were analysed by One-way of ANOVA followed by a post hoc test. Values are mean ± SEM with three different mice samples in each group. AS mice hippocampal tissues show significantly decreased levels of Arc and Egr3 (* P < 0.01) compared to the P25 WT group. Ube3a expression did not alter in the hippocampus of WT mice in response to eye-opening.

Article Snippet: Mouse monoclonal antibodies against Egr1 (sc-515830), Ube3a (sc-16689), Arc (sc-17839), and Egr3 (sc-390936) were purchased from Santa Cruz Biotechnology.

Techniques: Western Blot, Marker, Quantitation Assay, Expressing