cebpb antibody  (NSJ Bioreagents)

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

Journal: Cell Death Discovery

Article Title: Irisin inhibits adipogenic differentiation of bone marrow mesenchymal stem cells through the SIRT1/RANBP2/FTO signaling axis and protects against osteoporosis

doi: 10.1038/s41420-026-02976-5

Figure Lengend Snippet: A The effects of 10 μM, 25 μM, 50 μM, 75 μM, 100 μM, and 125 μM concentrations of irisin on the proliferation of BMMSCs were detected on days 12 h, 24 h, 36 h, and 48 h using the CCK-8 assay. B Flow chart of experiments to detect the osteogenic differentiation of BMMSCs. C Representative images of ARS staining during osteogenic (Osteo) differentiation in BMMSCs, and quantitative analysis of ARS staining. D Representative images of ALP staining during osteogenic (Osteo) differentiation in BMMSCs, and quantitative analysis of ALP staining. E After the addition of irisin, qRT-PCR was performed to analyze the mRNA levels of osteogenic (Osteo) related genes ( Runx2 , Col1 , and BMP2 ) in BMMSCs during the osteogenic differentiation. F Flow chart of experiments to detect the adipogenic differentiation of BMMSCs. G After the addition of irisin, qRT-PCR was performed to analyse the mRNA levels of adipogenic (Adipo) related genes ( Pparγ , C/ebpα , and C/ebpβ ) in BMMSCs during the adipogenic differentiation. H Representative images of Oil red O staining after adding irisin, and quantitative analysis of Oil red O staining. I After the addition of irisin, the levels of adipogenic (Adipo) related protein (PPARγ, C/EBPα, and C/EBPβ) in BMMSCs during the adipogenic differentiation were measured by western blotting. The values are mean ± SD of at least three independent experiments; n.s. p > 0.05, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: In brief, equal amounts of protein samples were separated on polyacrylamide gels and transferred to nitrocellulose membranes, which were closed for 1 h using 5% skimmed milk powder, and the membranes obtained were incubated with anti-FNDC5 antibody (#23995-1-AP, 1:1000, Proteintech, China), anti-PPARγ antibody (#7273, 1:1000, Santa Cruz Biotechnology, USA), anti-C/EBPα antibody (#2295, 1:1000, Cell Signaling Technology, USA), anti-C/EBPβ antibody (#3087, 1:1000, Cell Signaling Technology, USA), anti-SIRT1 antibody (#8469, 1:1000, Cell Signaling Technology, USA), anti-FTO antibody (#27226-1-AP, Proteintech, China), anti-RANBP2 antibody (#ab315458, 1:1000, Abcam, USA), and anti-GAPDH antibody (#10494-1, 1:1000, proteintech, China) were incubated at 4 °C overnight.

Techniques: CCK-8 Assay, Staining, Quantitative RT-PCR, Western Blot

Journal: Cell Death Discovery

Article Title: Irisin inhibits adipogenic differentiation of bone marrow mesenchymal stem cells through the SIRT1/RANBP2/FTO signaling axis and protects against osteoporosis

doi: 10.1038/s41420-026-02976-5

Figure Lengend Snippet: A Schematic workflow of the RNA-seq. B Volcanic maps of RNA-Seq in the PBS group and irisin group. C Heatmap of mRNA expression in the PBS group and irisin group, with high and low expression levels shown in red and blue respectively. D The mRNA levels of Serpina3n , Nos2 , Rdh9 , Serpina3m , and Sirt1 were detected by qRT-PCR. E qRT-PCR and western blotting confirmation of Sirt1 knockdown BMMSCs. F mRNA expressions of Pparγ , C/ebpα , and C/ebpβ in Sirt1 knockdown cells with or without irisin treatment. G Protein expressions of PPPARγ, C/EBPα, and C/EBPβ in Sirt1 knockdown cells with or without irisin treatment. H Representative images of Oil red O staining in Sirt1 knockdown cells with or without irisin treatment, and quantitative analysis of Oil red O staining. The values are mean ± SD of at least three independent experiments; n.s. p > 0.05, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: In brief, equal amounts of protein samples were separated on polyacrylamide gels and transferred to nitrocellulose membranes, which were closed for 1 h using 5% skimmed milk powder, and the membranes obtained were incubated with anti-FNDC5 antibody (#23995-1-AP, 1:1000, Proteintech, China), anti-PPARγ antibody (#7273, 1:1000, Santa Cruz Biotechnology, USA), anti-C/EBPα antibody (#2295, 1:1000, Cell Signaling Technology, USA), anti-C/EBPβ antibody (#3087, 1:1000, Cell Signaling Technology, USA), anti-SIRT1 antibody (#8469, 1:1000, Cell Signaling Technology, USA), anti-FTO antibody (#27226-1-AP, Proteintech, China), anti-RANBP2 antibody (#ab315458, 1:1000, Abcam, USA), and anti-GAPDH antibody (#10494-1, 1:1000, proteintech, China) were incubated at 4 °C overnight.

Techniques: RNA Sequencing, Expressing, Quantitative RT-PCR, Western Blot, Knockdown, Staining

Journal: Cell Death Discovery

Article Title: Irisin inhibits adipogenic differentiation of bone marrow mesenchymal stem cells through the SIRT1/RANBP2/FTO signaling axis and protects against osteoporosis

doi: 10.1038/s41420-026-02976-5

Figure Lengend Snippet: A Flowchart of SIRT1 destabilized FTO via RANBP2-mediated SUMOylation. B Fto mRNA expression in Sirt1 knockdown or Sirt1 -overexpressed cells was assayed by qRT-PCR. FTO protein expression in Sirt1 knockdown or Sirt1 -overexpressed cells was assayed by western blotting. C Fto mRNA expression in Sirt1 knockdown cells with or without irisin treatment was assayed by qRT-PCR. FTO protein expression in Sirt1 knockdown cells with or without irisin treatment was assayed by western blotting. D Immunoblotting of adipogenic related proteins (PPARγ, C/EBPα, and C/EBPβ) in Sirt1 -overexpressed cells with or without Fto overexpression. E Representative images of Oil red O staining in Sirt1 -overexpressed cells with or without Fto overexpression, and quantitative analysis of Oil red O staining. The values are mean ± SD of at least three independent experiments; n.s. p > 0.05, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: In brief, equal amounts of protein samples were separated on polyacrylamide gels and transferred to nitrocellulose membranes, which were closed for 1 h using 5% skimmed milk powder, and the membranes obtained were incubated with anti-FNDC5 antibody (#23995-1-AP, 1:1000, Proteintech, China), anti-PPARγ antibody (#7273, 1:1000, Santa Cruz Biotechnology, USA), anti-C/EBPα antibody (#2295, 1:1000, Cell Signaling Technology, USA), anti-C/EBPβ antibody (#3087, 1:1000, Cell Signaling Technology, USA), anti-SIRT1 antibody (#8469, 1:1000, Cell Signaling Technology, USA), anti-FTO antibody (#27226-1-AP, Proteintech, China), anti-RANBP2 antibody (#ab315458, 1:1000, Abcam, USA), and anti-GAPDH antibody (#10494-1, 1:1000, proteintech, China) were incubated at 4 °C overnight.

Techniques: Expressing, Knockdown, Quantitative RT-PCR, Western Blot, Over Expression, Staining

Journal: Cell Death Discovery

Article Title: Irisin inhibits adipogenic differentiation of bone marrow mesenchymal stem cells through the SIRT1/RANBP2/FTO signaling axis and protects against osteoporosis

doi: 10.1038/s41420-026-02976-5

Figure Lengend Snippet: A RANBP2 protein expression in Sirt1 knockdown cells with or without irisin treatment was assayed by western blotting. B Co-immunoprecipitation of SIRT1 with RANBP2 in BMMSCs with or without irisin treatment. C Colocalization of SIRT1 and RANBP2 after treatment with or without irisin in BMMSCs was exhibited in confocal immunofluorescent images. D Representative images of Oil red O staining in Sirt1 -overexpressed cells with or without Ranbp2 knockdown, and quantitative analysis of Oil red O staining. E mRNA expressions of Pparγ , C/ebpα , and C/ebpβ in Sirt1 -overexpressed cells with or without Ranbp2 knockdown. F Fto mRNA expression in Sirt1 -overexpressed cells with or without Ranbp2 knockdown was assayed by qRT-PCR. G FTO protein expression in Sirt1-overexpressed cells with or without Ranbp2 knockdown was assayed by western blotting. H Co-immunoprecipitation was used to detect the interactions of FTO with SIRT1 and RANBP2 proteins in irisin-treated or non-treated BMMSCs. Co-immunoprecipitation was used to detect the interactions of FTO with SUMO2/3 in irisin-treated or non-treated BMMSCs. The values are mean ± SD of at least three independent experiments; n.s. p > 0.05, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: In brief, equal amounts of protein samples were separated on polyacrylamide gels and transferred to nitrocellulose membranes, which were closed for 1 h using 5% skimmed milk powder, and the membranes obtained were incubated with anti-FNDC5 antibody (#23995-1-AP, 1:1000, Proteintech, China), anti-PPARγ antibody (#7273, 1:1000, Santa Cruz Biotechnology, USA), anti-C/EBPα antibody (#2295, 1:1000, Cell Signaling Technology, USA), anti-C/EBPβ antibody (#3087, 1:1000, Cell Signaling Technology, USA), anti-SIRT1 antibody (#8469, 1:1000, Cell Signaling Technology, USA), anti-FTO antibody (#27226-1-AP, Proteintech, China), anti-RANBP2 antibody (#ab315458, 1:1000, Abcam, USA), and anti-GAPDH antibody (#10494-1, 1:1000, proteintech, China) were incubated at 4 °C overnight.

Techniques: Expressing, Knockdown, Western Blot, Immunoprecipitation, Staining, Quantitative RT-PCR

Journal: Cell reports

Article Title: NAD + sensing by PARP7 regulates the C/EBPβ-dependent transcription program during adipogenesis

doi: 10.1016/j.celrep.2026.116929

Figure Lengend Snippet: (A) Schematic of experimental setup used for the genomic assays in 3T3-L1 cells. (B) Western blots showing the effects of Parp7 knockdown on the levels of total C/EBPβ and phosphorylated C/EBPβ (C/EBPβ-P) at day 1 of differentiation in 3T3-L1 cells. The cells were subjected to siRNA-mediated Parp7 knockdown 2 days before differentiation was induced using the MDI cocktail. (C) Browser tracks of genomic data at the Pparg gene. 3T3-L1 cells were subjected to control or Parp7 knockdown, followed by genomic assays: C/EBPβ and H3K27ac CUT&RUN, PARP7 ChIP-seq, and ATAC-seq. PARP7 ChIP-seq was performed after treatment with 400 nM RBN2397 to stabilize the PARP7 protein. (D) Heatmap showing gained, maintained, and depleted C/EBPβ CUT&RUN peaks upon siRNA-mediated knockdown of Parp7 in 3T3-L1 cells. (E) Metaplot of C/EBPβ CUT&RUN data centered at significant C/EBPβ peaks showing decreased C/EBPβ binding upon siRNA-mediated knockdown of Parp7 in 3T3-L1 cells. (F) Boxplot quantification of reads from C/EBPβ CUT&RUN peaks near 577 C/EBPβ target genes defined previously after siRNA-mediated knockdown of Parp7 in 3T3-L1 cells. The bars marked with different letters are significantly different from each other. Wilcox rank sum test, p < 4.652 × 10 −15 . (G) RNA-seq heatmap showing the regulation of 577 C/EBPβ target genes defined previously in 3T3-L1 cells with siRNA-mediated knockdown of Parp7 .

Article Snippet: C/EBPβ , Cell Signaling Technology , 3082; RRID:AB_2260365.

Techniques: Western Blot, Knockdown, Control, ChIP-sequencing, Binding Assay, RNA Sequencing

Journal: Cell reports

Article Title: NAD + sensing by PARP7 regulates the C/EBPβ-dependent transcription program during adipogenesis

doi: 10.1016/j.celrep.2026.116929

Figure Lengend Snippet: (A) Western blot showing a time course of PARP7 and C/EBPβ expression in 3T3-L1 cells differentiated with MDI cocktail. (B) Bar graphs showing the enrichment of C/EBPβ at the promoters of target genes Pparg , Atf3 , and Klf15 , as assayed by ChIP-qPCR, in 3T3-L1 cells with or without differentiation using MDI cocktail. Mean + SEM; n = 3. Asterisks indicate significant differences from control; ANOVA; * p < 0.05, ** p < 0.01, *** p < 0.005, and **** p < 0.001. (C) IP-western assays showing interactions between C/EBPβ and PARP7 in 3T3-L1 cells. 3T3-L1 cells with Dox-inducible ectopic expression of FLAG-C/EBPβ were subjected to FLAG IP. The IPs were subjected to western blotting for FLAG and PARP7. (D) Western blots of chromatin fractions from 3T3-L1 cells showing two p300-mediated histone modifications, H3K27ac and H2BK5ac, as well as H3K27me3, in bulk histones. PARP7 input confirms expected stabilization (2 nd lane from left) and knockdown (3 rd lane from left) of PARP7. (E) IP-western assays showing interactions between PARP7, p300, and C/EBPβ in 3T3-L1 cells. 3T3-L1 cells with Dox-inducible ectopic expression of FLAG-PARP7 were subjected to FLAG IP. The IPs were subjected to western blotting for FLAG, p300, and C/EBPβ. (F and G) Bar graphs showing the enrichment of (F) C/EBPβ or (G) p300 at the promoters of target genes Pparg , Atf3 , and Klf15 , as assayed by ChIP-qPCR, in 3T3-L1 cells differentiated for 24 h using MDI cocktail and subjected to siRNA-mediated depletion of Parp7 . Mean + SEM; n = 3. Asterisks indicate significant differences from control; Student’s t test; ** p < 0.01, *** p < 0.005, **** p < 0.001, and n.s., not significant. (H and I) Bar graphs showing the enrichment of (H) C/EBPβ and (I) H3K27ac at the promoters of target genes Pparg , Atf3 , and Klf15 , as assayed by ChIP-qPCR, in 3T3-L1 cells differentiated for 24 h using MDI cocktail and subjected to treatment with the p300 inhibitor A-485. Mean + SEM; n = 3. Asterisks indicate significant differences from control; Student’s t test; * p < 0.05, ** p < 0.01, and **** p < 0.001.

Article Snippet: C/EBPβ , Cell Signaling Technology , 3082; RRID:AB_2260365.

Techniques: Western Blot, Expressing, ChIP-qPCR, Control, Knockdown