cebpb antibody  (NSJ Bioreagents)

Journal: Molecular Biomedicine

Article Title: Curcumin alleviates metabolic dysfunction-associated steatotic liver disease via gut microbiota-dependent epigenetic alterations

doi: 10.1186/s43556-026-00606-1

Figure Lengend Snippet: CEBPB is regulated by AHR-mediated H3K27ac activation. a-b CEBPB protein abundance ( a ) and corresponding mRNA levels ( b ) in OA (0.6 mM)-induced HepG2 cells transfected with AHR overexpression plasmid ( AHR -OE) or empty vector (Vector) were examined by Western blotting and qRT-PCR, respectively ( n = 3). c-d The protein ( c ) and mRNA ( d ) levels of CEBPB in OA (0.6 mM)-induced Huh-7 cells transfected with siRNAs against AHR (si- AHR −1 and si- AHR −2) or negative control siRNA (si-NC) were determined by Western blotting and qRT-PCR, respectively ( n = 3). e A schematic diagram illustrating the predicted wild-type and mutant AHR-binding sites in the CEBPB promoter. f-g The transcriptional activity of wild-type and mutant CEBPB promoter constructs in response to AHR was determined by luciferase reporter assay, with Renilla luciferase activity serving as an input control ( n = 3). h ChIP-seq data from the Cistrome database (accession numbers: GSM1670897 , GSM2360939 ) show the enrichment of H3K27ac around the promoter region of CEBPB in human liver tissues, HepG2, and Huh-7 cells. i ChIP-seq data from the Cistrome database (accession numbers: ENCSR382MOM_1, ENCSR994YLZ_2, ENCSR994YLZ_1) reveal the enrichment of AHR around the promoter region of Cebpb in mouse liver tissues. j ChIP-qPCR validation of specific AHR occupancy at the Cebpb promoter region ( n = 3/group, two-way ANOVA). ChIP was conducted with AHR antibody and non-specific IgG control. k-l ChIP assays were performed to determine whether AHR modulates H3K27ac enrichment at the CEBPB regulatory locus in HepG2 ( k ) and Huh-7 ( l ) cells. m A schematic diagram depicting an interactive model that AHR transcriptionally activates CEBPB expression. Results were presented as mean ± SD. P values were calculated with unpaired two-sided Student’s t -test in ( b ) and f-g , one-way ANOVA followed by Tukey’s test in d . * P < 0.05, ** P < 0.01, *** P < 0.001, ns: not significant

Article Snippet: The primary antibodies used were as follows: AHR (67785–1-Ig, Proteintech, China), CEBPB (23431–1-AP, Proteintech, China), ACC1 (21923–1-AP, Proteintech, China), CD36 (18836–1-AP, Proteintech, China), SCD1 (28678–1-AP, Proteintech, China), phosphorylated PI3K (p-PI3K, 138364, Abcam, USA), PI3K (4257, CST, USA), phosphorylated AKT (p-AKT, 66444–1-Ig, Proteintech, China), AKT (10176–2-AP, Proteintech, China), and β-actin (66009–1-Ig, Proteintech, China).

Techniques: Activation Assay, Quantitative Proteomics, Transfection, Over Expression, Plasmid Preparation, Western Blot, Quantitative RT-PCR, Negative Control, Mutagenesis, Binding Assay, Activity Assay, Construct, Luciferase, Reporter Assay, Control, ChIP-sequencing, ChIP-qPCR, Biomarker Discovery, Expressing

Journal: Molecular Biomedicine

Article Title: Curcumin alleviates metabolic dysfunction-associated steatotic liver disease via gut microbiota-dependent epigenetic alterations

doi: 10.1186/s43556-026-00606-1

Figure Lengend Snippet: C. sensu regulated lipid accumulation in hepatocytes in a CEBPB-dependent manner. a-d Representative BODIPY 493/503 fluorescence staining of lipid droplet accumulation and quantitative analysis in HepG2 and Huh-7 cells treated with C. sensu or control and transfected with siRNAs against CEBPB (si- CEBPB ) or negative control siRNA (si-NC) with or without OA (0.6 mM) treatment (scale bar, 100 μm, n = 3). e–f Quantification of intracellular TG content in HepG2 and Huh-7 cells treated with C. sensu or control transfected with si- CEBPB or si-NC with or without OA treatment (0.6 mM, n = 3). g-j Representative ORO staining of lipid droplet accumulation and quantitative analysis in HepG2 and Huh-7 cells treated with C. sensu or Control and transfected with si- CEBPB or si-NC, with or without OA (0.6 mM) treatment (scale bar, 100 μm, n = 3). k The levels of lipid transport- and synthesis-related proteins ( ACC1 , CD36 , and SCD1 ) in C. sensu - or control-treated HepG2 and Huh-7 cells transfected with si- CEBPB or si-NC with or without OA treatment (0.6 mM) were assessed by Western blotting ( n = 3). Results were presented as mean ± SD from three independent experiments. P values were calculated with one-way ANOVA followed by Tukey’s test. The abbreviation C. sensu represents Clostridium_sensu_stricto_1 . * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: The primary antibodies used were as follows: AHR (67785–1-Ig, Proteintech, China), CEBPB (23431–1-AP, Proteintech, China), ACC1 (21923–1-AP, Proteintech, China), CD36 (18836–1-AP, Proteintech, China), SCD1 (28678–1-AP, Proteintech, China), phosphorylated PI3K (p-PI3K, 138364, Abcam, USA), PI3K (4257, CST, USA), phosphorylated AKT (p-AKT, 66444–1-Ig, Proteintech, China), AKT (10176–2-AP, Proteintech, China), and β-actin (66009–1-Ig, Proteintech, China).

Techniques: Fluorescence, Staining, Analysis, Control, Transfection, Negative Control, Western Blot

Journal: Molecular Biomedicine

Article Title: Curcumin alleviates metabolic dysfunction-associated steatotic liver disease via gut microbiota-dependent epigenetic alterations

doi: 10.1186/s43556-026-00606-1

Figure Lengend Snippet: Reduced KYN plays a protective role in Cur-regulated lipid steatosis in MASLD. a Representative morphology images of livers from rats fed ND, HFD, HFD + Cur, and HFD + Cur + KYN (scale bar, 1 cm, n = 6). b Histopathological section of liver tissue by hematoxylin and eosin staining (scale bar, 50 μm, n = 6). c Oil Red O (ORO) staining of the livers from the indicated groups (scale bar, 50 μm, n = 6). d Transmission electron microscopy (TEM) of hepatocytes in ND, HFD, HFD + Cur, and HFD + Cur + KYN groups at 3600 × (upper, scale bar, 2 μm) and 8500 × (lower, scale bar, 1 μm) magnification ( n = 6). e Body weight of ND, HFD, HFD + Cur, and HFD + Cur + KYN rats ( n = 6). f Liver index (ratio of liver weight to body weight) was calculated for ND, HFD, HFD + Cur, and HFD + Cur + KYN rats ( n = 6). g MASLD activity score (MAS) in the indicated groups ( n = 6). h-m Serum TG ( h ), TC ( i ), AST ( j ), ALT ( k ), LDL-C ( l ), and HDL-C ( m ) levels in the indicated groups ( n = 6). n . Graphical abstract. A comprehensive multi-omics study integrating 16S microbiomics, metabolomics, transcriptomics, and H3K27ac epigenomics was conducted to elucidate the mechanisms of Cur’s anti-MASLD effect. Oral administration of Cur demonstrated a beneficial impact on MASLD by modulating the gut microbiota, notably suppressing the pathogenic bacterium C. sensu , which subsequently led to a decrease in KYN accumulation. Mechanistically, gut microbiota-derived KYN binds to AHR, recruiting the epigenetic histone modification H3K27ac to specifically regulate the key gene Cebpb , further activating PI3K-AKT pathway to aggravate hepatic steatosis. Collectively, this study reveals a novel gut-liver axis in MASLD pathogenesis, demonstrating that Cur alleviates hepatic steatosis by targeting the microbiota-KYN-AHR-H3K27ac- Cebpb epigenetic axis, thereby positioning both Cur and KYN modulation as promising therapeutic strategies for MASLD intervention (Created with BioRender). All data are represented as the mean ± SD. P values were calculated by two-way ANOVA followed by Bonferroni post-test in ( e ), others with one-way ANOVA followed by Tukey’s test. * P < 0.05, ** P < 0.01, *** P < 0.001. ALT: alanine aminotransferase, AST: aspartate aminotransferase, TG: triglycerides, TC: total cholesterol, HDL-C: high-density lipoprotein cholesterol, LDL-C: low-density lipoprotein cholesterol

Article Snippet: The primary antibodies used were as follows: AHR (67785–1-Ig, Proteintech, China), CEBPB (23431–1-AP, Proteintech, China), ACC1 (21923–1-AP, Proteintech, China), CD36 (18836–1-AP, Proteintech, China), SCD1 (28678–1-AP, Proteintech, China), phosphorylated PI3K (p-PI3K, 138364, Abcam, USA), PI3K (4257, CST, USA), phosphorylated AKT (p-AKT, 66444–1-Ig, Proteintech, China), AKT (10176–2-AP, Proteintech, China), and β-actin (66009–1-Ig, Proteintech, China).

Techniques: Morphology, Tissue, H&E Stain, Staining, Transmission Assay, Electron Microscopy, Activity Assay, Biomarker Discovery, Transcriptomics, Derivative Assay, Modification, RNA Sequencing, ChIP-sequencing

Journal: iMetaOmics

Article Title: Integrin α5β1‐mediated multicellular crosstalk in the tumor microenvironment drives bladder cancer progression and reveals targetable vulnerabilities

doi: 10.1002/imo2.70102

Figure Lengend Snippet: Single‐cell expression profiling identifies and characterizes malignant epithelial clusters in BCa. (A) Schematic overview of the study design, illustrating the sampling and analytical workflow. (B, C) UMAP visualization of 178,633 cells derived from 24 samples, encompassing NAT, GC, Low_grade, and High_grade tissues, identifying seven major cell types. (D) Expression profiles of established markers across the designated cell clusters. (E) Box plots illustrating the proportions of the seven major cell types across different pathological stages. (F) Re‐clustering of epithelial cells revealing distinct heterogeneity across NAT, GC, Low_Grade, and High_Grade tissues. (G) CNV score highlighting genomic instability across epithelial subclusters. (H) Pathway enrichment scores in different epithelial cell clusters (labeled as g0, g1, etc.), as determined by GSVA. (I) Violin plots illustrating the upregulation of malignancy‐associated genes in epithelial cells during BCa progression. (J) Dot plots displaying the expression levels of malignancy‐associated genes across re‐clustered epithelial cell groups. (K) Pseudotime developmental trajectory of epithelial cells inferred using Monocle2 and CytoTRACE2. (L) Heatmap displaying the dynamic expression patterns of malignancy‐associated genes along the pseudotime trajectory. (M) Stemness scores for different epithelial cell clusters calculated with AddModuleScore. (N) GO enrichment analysis highlighting key biological processes associated with Cluster 1_SDC1. (O) Validation of SDC1 knockdown efficiency by Western blot and qRT‐PCR in UMUC3 cells transfected with control or SDC1 siRNA. (P) CCK8 assay assessing the effect of SDC1 knockdown on UMUC3 cell proliferation. (Q) Representative images of colony formation in UMUC3 cells after SDC1 knockdown, showing colony size. Scale bar = 100 μm. Statistical analysis of the colony formation assay is shown on the right ( n ≥ 10). (R) CellChat analysis illustrating altered intercellular signaling strengths received by epithelial cells in high‐grade tumors. Data are mean ± SD. Analyzed by Student's t ‐test (O, Q) or two‐way ANOVA with Dunnett's test (P). ** p < 0.01, *** p < 0.001 versus NC. BCa, bladder cancer; CCK8, Cell Counting Kit‐8; CNV, copy number variation; Comun. Prob., communication probability; GC, cystitis glandularis; GO, Gene Ontology; GSVA, Gene Set Variation Analysis; NAT, normal adjacent tissue; NC, negative control; pct.exp, percentage of expressed cells; qRT‐PCR, quantitative real‐time PCR; SDC1, syndecan 1; siRNA, small interfering RNA; UMAP, Uniform Manifold Approximation and Projection.

Article Snippet: Monocytic MDSCs (CD11b+CD33+HLA‐DR^low) were sorted by flow cytometry (BD Biosciences) and transfected with CEBPB siRNA (HY‐RS02446, MCE).

Techniques: Single Cell, Expressing, Profiling, Sampling, Derivative Assay, Labeling, Analysis, Biomarker Discovery, Knockdown, Western Blot, Quantitative RT-PCR, Transfection, Control, CCK-8 Assay, Colony Assay, Cell Counting, Tissue, Negative Control, Real-time Polymerase Chain Reaction, Small Interfering RNA

Journal: iMetaOmics

Article Title: Integrin α5β1‐mediated multicellular crosstalk in the tumor microenvironment drives bladder cancer progression and reveals targetable vulnerabilities

doi: 10.1002/imo2.70102

Figure Lengend Snippet: Transcriptional characterization and identification of myeloid‐derived suppressor cell (MDSC) subsets in high‐grade BCa. (A) UMAP plot illustrating the classification of myeloid cell subtypes. (B) Dot plots displaying the expression of common marker genes across each myeloid cell subset. (C) Bar plots illustrating the total cell numbers of each myeloid cell subset across various sample types. (D) Dot plots depicting Spearman correlation between CAFs and myeloid subtype infiltration in the TCGA‐BLCA cohort. The size of each dot is proportional to the absolute magnitude of the correlation coefficient, and the color intensity indicates the correlation value (red for positive, blue for negative). (E) Mountain plot illustrating the enrichment scores of selected Hallmark gene sets across different myeloid cell subtypes. (F) GO enrichment analysis highlighting BP and CC pathways based on highly variable genes in the Mono_TYMP subset. Bar length denotes enrichment significance (−log 10 p value), while the yellow line represents the percent of geneRatio. (G) Pseudotime trajectory inferring the developmental evolution of myeloid cells. (H) Expression profiles of immune suppression‐related molecules across different myeloid cell subtypes. (I) Estimation of M‐MDSC‐related gene set scores across myeloid cell subtypes using AddModuleScore. (J) M‐MDSC scoring in the tumor progression of LUAD ( GSE189357 ) and BCa. p values were calculated via two‐sided Wilcoxon rank‐sum test. (K) CellPhoneDB analysis of ligand–receptor interactions in coinhibitory signaling pathways between different myeloid cell subtypes and T/NK cells. (L) Bubble plot illustrating signaling pathway interactions from different myeloid cell subtypes to endothelial cells. (M) mIF staining visualizing the spatial proximity between NAMPT+ cells and ITGB1+ cells. Quantification is shown on the right. Data are presented as mean ± SD. p values were calculated via Student's t ‐test. Scale bar = 10 μm. (N) Representative mIF images showing colocalization of VEGF‐expressing MDSCs and endothelial cells (CD31). Scale bar = 10 μm. (O) Mountain plot showing the activity of the STAT3 and CEBPB regulons. (P) Regulatory network plot illustrating the transcription factors (STAT3 and CEBPB) and their target genes. Node size is proportional to the node degree (number of connections). Node and edge colors represent the specific transcription factor regulons (STAT3 network in red, CEBPB network in blue). (Q) Spearman correlation analysis showing the association between the expression of CEBPB and NAMPT , as well as the angiogenesis‐related gene RHOB , in the TCGA‐BLCA cohort. (R) Schematic diagram of the regulatory network. Black arrows indicate transcriptional activation of target genes by STAT3 and CEBPB. Red upward arrows denote the upregulation of these genes. * p < 0.05; ** p < 0.01; *** p < 0.001. BP, biological process; CC, cellular component; LUAD, lung adenocarcinoma; MDSC, myeloid‐derived suppressor cell; M‐MDSC, monocytic MDSC.

Article Snippet: Monocytic MDSCs (CD11b+CD33+HLA‐DR^low) were sorted by flow cytometry (BD Biosciences) and transfected with CEBPB siRNA (HY‐RS02446, MCE).

Techniques: Derivative Assay, Expressing, Marker, Analysis, Protein-Protein interactions, Staining, Activity Assay, Activation Assay

Journal: Journal of Hepatocellular Carcinoma

Article Title: C/EBP-β Mediates the Reversal of Sorafenib Resistance by Tunicamycin in Hepatocellular Carcinoma

doi: 10.2147/JHC.S587037

Figure Lengend Snippet: C/EBP-β is downregulated in sorafenib-resistant hepatocellular carcinoma cells. ( A ) The inhibition ratio (%) of parental (HepG2, Huh7) and sorafenib-resistant (HepG2-R, Huh7-R) HCC cells was measured by CCK-8 assay after treatment with a gradient of sorafenib concentrations for 48 h. The calculated IC 50 values are indicated on the dose-response curves. Data are presented as mean ± SD (n=3). ( B ) The relative mRNA and protein expression levels of C/EBP-β in parental and sorafenib-resistant cell lines were determined by RT-qPCR (bar graphs) and Western blot (gel images), respectively. GAPDH was used as an internal control. Data in the bar graphs are presented as mean ± SD from three independent experiments. ** P < 0.01 compared to the corresponding parental cells.

Article Snippet: The following primary antibodies were used for Western blot and immunohistochemistry: C/EBP-β (A0711, Abclonal, China), p53 (ET1601-13, HUABIO, China), Bax (ET1603-34, HUABIO, China), Bcl-2 (ET1702-53, HUABIO, China), Cleaved Caspase-3 (ET1602-47, HUABIO, China), GAPDH (K200102M, Solarbio, China), and Ki-67 (PT0321R, Immunoway, China).

Techniques: Inhibition, CCK-8 Assay, Expressing, Quantitative RT-PCR, Western Blot, Control

Journal: Journal of Hepatocellular Carcinoma

Article Title: C/EBP-β Mediates the Reversal of Sorafenib Resistance by Tunicamycin in Hepatocellular Carcinoma

doi: 10.2147/JHC.S587037

Figure Lengend Snippet: Tunicamycin (TM) reverses sorafenib resistance by upregulating C/EBP-β. ( A ) Western blot analysis of C/EBP-β protein levels in parental Huh7 and resistant Huh7-R cells treated with increasing concentrations of TM (0, 2, 4, 8 μM) for 48 h. ( B ) RT-qPCR analysis of relative C/EBP-β mRNA levels in cells treated as in ( A ). ( C ) Cell viability of Huh7-R cells was measured by CCK-8 assay. Cells were treated with 8 μM sorafenib alone or in combination with TM (2, 4, 8 μM) for the indicated durations. Parental Huh7 cells treated with 8 μM sorafenib served as a sensitive control. ( D ) Clonogenic potential of cells treated as in ( C ). Representative images (left) and quantification (right) are shown. ( E ) Western blot validation of C/EBP-β overexpression (C/EBP-β-oe) and knockdown (shC/EBP-β) in Huh7-R cells under various treatment conditions. ( F ) Cell viability (CCK-8) of genetically modified or unmodified Huh7-R cells treated with sorafenib (8 μM) with or without TM (4 μM) for 48 h. ( G ) Cell viability (CCK-8) of HepG2-R cells treated as in ( F ). ( H ) Clonogenic potential of Huh7-R cells treated as in ( F ). ( I ) Clonogenic potential of HepG2-R cells treated as in ( F ). The “+” and “-” signs indicate the presence or absence of the specified treatment, respectively. All quantitative data are presented as mean ± SD from three independent experiments. * P < 0.05, ** P < 0.01, and *** P < 0.001 compared between indicated groups.

Article Snippet: The following primary antibodies were used for Western blot and immunohistochemistry: C/EBP-β (A0711, Abclonal, China), p53 (ET1601-13, HUABIO, China), Bax (ET1603-34, HUABIO, China), Bcl-2 (ET1702-53, HUABIO, China), Cleaved Caspase-3 (ET1602-47, HUABIO, China), GAPDH (K200102M, Solarbio, China), and Ki-67 (PT0321R, Immunoway, China).

Techniques: Western Blot, Quantitative RT-PCR, CCK-8 Assay, Control, Biomarker Discovery, Over Expression, Knockdown, Genetically Modified

Journal: Journal of Hepatocellular Carcinoma

Article Title: C/EBP-β Mediates the Reversal of Sorafenib Resistance by Tunicamycin in Hepatocellular Carcinoma

doi: 10.2147/JHC.S587037

Figure Lengend Snippet: TM and C/EBP-β overexpression cooperatively suppress sorafenib-resistant tumor growth in vivo. Nude mice were treated with sorafenib (15 mg/kg/day) alone (control), sorafenib plus tunicamycin (TM, 0.04 mg/kg/day), sorafenib plus C/EBP-β overexpression (C/EBP-β-oe), or a combination of sorafenib, TM, and C/EBP-β-oe. ( A – D ) Subcutaneous xenograft model using Huh7-R cells. ( A ) Representative images of excised tumors from each treatment group on day 38. ( B ) Tumor growth curves measured over a 38-day treatment period. Data are presented as mean ± SD (n=6 mice per group). **P< 0.01 and ***P< 0.001 compared with the sorafenib-only group. ## P< 0.01 compared between indicated groups. ( C ) H&E staining of tumor sections showing tumor morphology and cell density. Scale bars, 50 μm. ( D ) Immunohistochemical staining for C/EBP-β and proliferation marker Ki-67 in tumor tissues. Scale bars, 50 μm. ( E and F ) Orthotopic xenograft model of Huh7-R cells. ( E ) Representative images of the liver illustrating intrahepatic tumor burden at the experimental endpoint. ( F ) Representative H&E staining of liver sections from the orthotopic model, with insets providing magnified views of tumor foci. Scale bars: 100 μm (main) and 50 μm (inset). The “+” and “-” signs indicate the presence or absence of the specified treatment, respectively.

Article Snippet: The following primary antibodies were used for Western blot and immunohistochemistry: C/EBP-β (A0711, Abclonal, China), p53 (ET1601-13, HUABIO, China), Bax (ET1603-34, HUABIO, China), Bcl-2 (ET1702-53, HUABIO, China), Cleaved Caspase-3 (ET1602-47, HUABIO, China), GAPDH (K200102M, Solarbio, China), and Ki-67 (PT0321R, Immunoway, China).

Techniques: Over Expression, In Vivo, Control, Staining, Immunohistochemical staining, Marker

Journal: Journal of Hepatocellular Carcinoma

Article Title: C/EBP-β Mediates the Reversal of Sorafenib Resistance by Tunicamycin in Hepatocellular Carcinoma

doi: 10.2147/JHC.S587037

Figure Lengend Snippet: The C/EBP-β axis overcomes sorafenib resistance by inducing ROS production and apoptosis in Huh7-R cells. Huh7-R cells were treated for 48 h with sorafenib (8 μM) alone or in combination with C/EBP-β overexpression (C/EBP-β-oe), tunicamycin (TM, 4 μM), TM plus shC/EBP-β, or TM plus NAC (5 mM). Hydrogen peroxide (H 2 O 2 , 2 mM) was included as a positive control for ROS generation. ( A ) Intracellular ROS levels were detected using DCFDA staining and visualized using fluorescence microscopy. Scale bar, 200 μm. ( B ) Quantification of the relative fluorescence intensity from ( A ). ( C ) Apoptosis was detected using the TUNEL assay, with TUNEL-positive nuclei stained green and all nuclei counterstained with DAPI (blue). Scale bar, 100 μm. ( D ) Percentage of TUNEL-positive cells from ( C ). ( E ) Western blot analysis of C/EBP-β, p53, Bax, Bcl-2, and cleaved caspase-3 protein levels. GAPDH was used as an internal control. The “+” and “-” signs indicate the presence or absence of the specified treatment, respectively. All quantitative data are presented as mean ± SD from three independent experiments. * P < 0.05, ** P < 0.01, and *** P < 0.001 compared between indicated groups.

Article Snippet: The following primary antibodies were used for Western blot and immunohistochemistry: C/EBP-β (A0711, Abclonal, China), p53 (ET1601-13, HUABIO, China), Bax (ET1603-34, HUABIO, China), Bcl-2 (ET1702-53, HUABIO, China), Cleaved Caspase-3 (ET1602-47, HUABIO, China), GAPDH (K200102M, Solarbio, China), and Ki-67 (PT0321R, Immunoway, China).

Techniques: Over Expression, Positive Control, Staining, Fluorescence, Microscopy, TUNEL Assay, Western Blot, Control

Journal: Journal of Hepatocellular Carcinoma

Article Title: C/EBP-β Mediates the Reversal of Sorafenib Resistance by Tunicamycin in Hepatocellular Carcinoma

doi: 10.2147/JHC.S587037

Figure Lengend Snippet: Integrated bioinformatic analyses define the C/EBP-β-dependent transcriptomic mechanism of tunicamycin in HCC. ( A ) Representative Gene Set Enrichment Analysis (GSEA) plots from the TCGA-LIHC dataset showing positive enrichment of apoptosis (TWEAK signaling) and ER stress (ATF4 activation) pathways in tumors with high C/EBP-β expression. ( B ) Venn diagram illustrating the identification of 185 intersecting genes between potential Tunicamycin (TM) targets (from SwissTarget, Pharmmapper, and DrugBank) and known hepatocellular carcinoma (HCC) disease genes (from GeneCards and OMIM). ( C ) GO (Biological Process) and KEGG pathway enrichment analysis of the 185 intersecting genes identified in (B). The bubble plot displays top enriched terms, with bubble size corresponding to gene count and color intensity reflecting the adjusted P -value. ( D ) GO enrichment analysis (Biological Process) of the 13 core genes at the intersection of TM’s potential targets and C/EBP-β-regulated differentially expressed genes. The bar chart highlights “intrinsic apoptotic signaling pathway in response to ER stress” and “multiple cell differentiation-related pathways” as the key convergent mechanisms.

Article Snippet: The following primary antibodies were used for Western blot and immunohistochemistry: C/EBP-β (A0711, Abclonal, China), p53 (ET1601-13, HUABIO, China), Bax (ET1603-34, HUABIO, China), Bcl-2 (ET1702-53, HUABIO, China), Cleaved Caspase-3 (ET1602-47, HUABIO, China), GAPDH (K200102M, Solarbio, China), and Ki-67 (PT0321R, Immunoway, China).

Techniques: Activation Assay, Expressing, Cell Differentiation

Journal: Cancer Communications

Article Title: CEBPB-Regulated Gastric Cell Plasticity Promotes Liver Metastasis of Gastric Cancer

doi: 10.34133/cancomm.0016

Figure Lengend Snippet: CEBPB regulates gastric plasticity through its global enhancer-binding function. (A) H3K27ac modification levels, chromosomal accessibility (defined by ATAC-seq signals), and the number of CEBPB-binding sites across the genome in the NCI-N87 ( n = 2). (B) CEBPB Cut&Tag, H3K27ac Cut&Tag, and ATAC-seq peak signal levels flanking ±5 kb of the CEBPB binding sites (indicated by the black box) in NCI-N87 ( n = 2). (C) Venn diagrams showing the overlap between putative enhancers and CEBPB binding sites. (D) Gene set enrichment analysis of malignant and nonmalignant cell clusters using the top 200 most active CEBPB-binding enhancer genes derived from NCI-N87 H3K27ac Cut&Tag data. (E) Gene set enrichment analysis of transcriptome profiles of GC cell lines derived from GCLM (red) and primary GC tissues (blue) using the top 200 most active CEBPB-binding enhancer genes derived from NCI-N87 H3K27ac Cut&Tag data. (F) Gene set enrichment analysis of transcriptome profiles of CTCs using the top 200 most active CEBPB-binding enhancer genes derived from NCI-N87 H3K27ac Cut&Tag data. (G to I) Expression levels of liver-specific genes ( SLC27A5 , F12 , and DECR2 ) in malignant and nonmalignant cells (G), CTCs (H), and GC cell lines (I). (J) ATAC-seq, H3K27ac Cut&Tag, and CEBPB Cut&Tag signal levels, along with putative enhancer and CEBPB binding regions, in the gene regions of SLC27A5 , F12 , and DECR2 in NCI-N87 ( n = 2). Statistical significance, * P < 0.05, ** P < 0.01, *** P < 0.001. Abbreviations: ATAC-seq, assay for transposase-accessible chromatin using sequencing; CEBPB, CCAAT enhancer-binding protein beta; CPM, counts per million; Cut&Tag, cleavage under targets and tagmentation; CTC, circulating tumor cell; CTC-LM, CTC associated with liver metastasis; CTC-N, CTC not associated with liver metastasis; DECR2, 2,4-dienoyl-CoA reductase 2; F12, coagulation factor XII; GC, gastric cancer; GCLM, gastric cancer liver metastasis; Mag, malignant; Rep, replicate; SLC27A5, solute carrier family 27 member 5; TPM, transcripts per million.

Article Snippet: The cells were incubated overnight with primary antibodies anti-CEBPB (Cat. #23431-1-AP, Proteintech), anti-H3K27ac (Cat. #13-0059, EpiCypher), and IgG negative control (Cat. #13-0047, EpiCypher).

Techniques: Binding Assay, Modification, Derivative Assay, Expressing, Sequencing, Coagulation

Journal: Cancer Communications

Article Title: CEBPB-Regulated Gastric Cell Plasticity Promotes Liver Metastasis of Gastric Cancer

doi: 10.34133/cancomm.0016

Figure Lengend Snippet: Cebpb-regulated gastric plasticity evades CD8 + T cell surveillance by CD155–TIGIT interaction in vivo. (A) Schematic of in vivo mouse experiments to determine the immune-regulatory role of Cebpb ( n = 5 for each group). The healthy control group denoted the group without cancer cell transplantation, and the NC group denoted the group transplanted with MFC. (B) Tumor volume of liver metastases inferred from in vivo bioluminescence imaging results performed every 7 days from day 3 to day 50 (left) since cell transplantation, and liver metastases visualized using a bioluminescence imaging system on day 50 (right). (C) Representative H&E staining and immunohistochemistry results for Cebpb in liver metastasis tissues from the control group, MFC Cebpb-OE group, and Cebpb-OE plus anti-TIGIT mAbs treatment group. Arrows denote the representative areas of GC liver metastasis within the hepatic tissue. (D) Levels of IFN-γ, TNF-α, and the proportion of CD8 + T cells and NK cells among CD45 + immune cells in the healthy control group (green), MFC control group (gray), Cebpb-OE group (red), and Cebpb-OE plus anti-TIGIT mAbs treatment group (blue). (E) Multiplex immunofluorescence showing CD8 (green), IFN-γ (red), and nuclei (blue) in the MFC control group, Cebpb-OE group, and Cebpb-OE plus anti-TIGIT mAbs treatment group. Data were presented as the mean ± standard deviation. Statistical significance, * P < 0.05, ** P < 0.01, *** P < 0.001. Abbreviations: Cebpb, CCAAT enhancer binding protein beta; H&E, hematoxylin and eosin; IFN-γ, interferon-gamma; mAb, monoclonal antibody; NC, negative control; OE, overexpression; TNF-α, tumor necrosis factor alpha; TIGIT, T cell immunoreceptor with Ig and ITIM domains.

Article Snippet: The cells were incubated overnight with primary antibodies anti-CEBPB (Cat. #23431-1-AP, Proteintech), anti-H3K27ac (Cat. #13-0059, EpiCypher), and IgG negative control (Cat. #13-0047, EpiCypher).

Techniques: In Vivo, Control, Transplantation Assay, Imaging, Staining, Immunohistochemistry, Multiplex Assay, Immunofluorescence, Standard Deviation, Binding Assay, Negative Control, Over Expression