anti cbx3  (Proteintech)

Journal: Oncogene

Article Title: CBX3 promotes multidrug resistance by suppressing ferroptosis in colorectal carcinoma via the CUL3/NRF2/GPX2 axis

doi: 10.1038/s41388-025-03337-9

Figure Lengend Snippet: A Association of CBX3 with overall survival and disease-free survival in CRC from the TCGA database. B Association of CBX3 with disease-free survival in CRC after chemotherapy. C Analysis of CBX3 expression in CRC cases after chemotherapy, divided into non-Response and Response groups. D Representative immunohistochemical images of CRC tissue stained with anti-CBX3 and anti-Ki67 antibodies (low-power view 10×, high-power view 40×, scale bars = 50 μm). Spearman’s correlation analysis of CBX3 and Ki67 expression was shown. E Western blotting of various CRC cells. β-actin serves as an internal reference. Stable CBX3-overexpressed cells were SW480-CBX3, compared with SW480 WT cells (Con); whereas siCBX3 was transfected into HCT116 cells, compared with siNC. F Cell counting assays in SW480-Con/CBX3 and HCT116 siNC/siCBX3 cells with Irinotecan (1 μmol/L) and Oxaliplatin (0.5 μmol/L) treatments. Data were presented as the means ± SEM ( N = 4). G Analysis of drug sensitivity in various SW480 and HCT116 cell lines after 48 h of treatment with the indicated drug concentrations. Data were presented as the means ± SEM ( N = 5). * P < 0.05, ** P < 0.01, compared to Con group with solvent treatment. Iri Irinotecan, Oxa Oxaliplatin.

Article Snippet: A pCMV-based plasmid encoding human CBX3 and a pcDNA3-based plasmid encoding human NFE2L2 were obtained from the nonprofit plasmid repository, Addgene (Cambridge, MA, USA).

Techniques: Expressing, Immunohistochemical staining, Staining, Western Blot, Transfection, Cell Counting, Solvent

Journal: Oncogene

Article Title: CBX3 promotes multidrug resistance by suppressing ferroptosis in colorectal carcinoma via the CUL3/NRF2/GPX2 axis

doi: 10.1038/s41388-025-03337-9

Figure Lengend Snippet: A Western blotting assays on caspase-3 and cleaved caspase-3 in SW480-Con/CBX3 cells following chemotherapy. β-Actin served as an internal reference. B Cell counting assays in SW480-Con/CBX3 cells with Z-VAD, Necrostain, and various chemotherapies. Data were presented as the means ± SEM ( N = 4). C KEGG pathway enrichment analysis for CBX3-related gene expression in CRC cases from the TCGA database, divided into high and low CBX3 expression groups. D Cell counting assays in SW480-Con/CBX3 cells treated with ferroptosis inhibitor Fer-1 and various chemotherapies. Data were presented as the means ± SEM ( N = 4). E MTT assays to detect cell viability in various SW480 or HCT116 cells after a 48-h treatment with the ferroptosis activators RSL3 and FIN56. Data were presented as the means ± SEM ( N = 5). F GSH/GSSG assay in SW480-Con/CBX3 cells following chemotherapy. F GSH/GSSG assay in SW480-Con/CBX3 cells following chemotherapy. G Lipid peroxidation assays and the cellular location in CRC cells after chemotherapy. SW480-Con/CBX3 cells were treated with DMSO or drugs for 8 h and stained with C11 BODIPY (oxidized lipid was stained green) and then detected by both flow cytometry and confocal microscopy. ERTracker Blue-White was used to detect the location of lipid oxidation by confocal microscopy (high-power view 400×, scale bar = 50 μm) ( N = 3). H Mitochondrial electron microscopy assays in SW480-Con/CBX3 cells following chemotherapy (left: lower power view, 10,000×, bar = 1 μm; right: high-power view, 30,000×, bar = 500 nm; arrows: damaged mitochondria). A – G Data were presented as the means ± SEM ( N = 3). Con SW480-Con cells, CBX3 SW480-CBX3 cells, NC Solvent treatment, Iri Irinotecan, Oxa Oxaliplatin. * P < 0.05, ** P < 0.01, compared to corresponding groups.

Article Snippet: A pCMV-based plasmid encoding human CBX3 and a pcDNA3-based plasmid encoding human NFE2L2 were obtained from the nonprofit plasmid repository, Addgene (Cambridge, MA, USA).

Techniques: Western Blot, Cell Counting, Gene Expression, Expressing, GSSG Assay, Staining, Flow Cytometry, Confocal Microscopy, Electron Microscopy, Solvent

Journal: Oncogene

Article Title: CBX3 promotes multidrug resistance by suppressing ferroptosis in colorectal carcinoma via the CUL3/NRF2/GPX2 axis

doi: 10.1038/s41388-025-03337-9

Figure Lengend Snippet: A RNA-sequence analysis of SW480-Con/CBX3 cells. NRF2-signaling pathway was enriched in oncogenic signatures. B Association of NRF2 with overall survival in CRC from the TCGA database. The expression of the NRF2-related pathway was analyzed in CRC cases using the CPTAC database. C Representative immunohistochemical images of CRC tissue stained with anti-CBX3 and anti-NRF2 antibodies (low-power view 10×, high-power view 40×, scale bars = 50 μm). Spearman’s correlation analysis of CBX3 and NRF2 expression is shown. D Western blotting of CBX3 and NRF2 in various CRC cells. E Western blotting of NRF2 in CBX3-overexpressed HEK-293T and SW480 cells. β-actin serves as an internal reference. F Western blotting of NRF2 in siCBX3-transfected HCT116 cells. β-actin serves as an internal reference. G Analysis of drug sensitivity in SW480-CBX3 cells with and without siNRF2 after treatment with the indicated drug concentrations. H GSH/GSSG and MDA assays in SW480-Con/CBX3 cells following chemotherapy. B – H Con: SW480-Con cells, CBX3: SW480-CBX3 cells. Data were presented as the means ± SEM ( N = 3). * P < 0.05, ** P < 0.01, compared to corresponding groups. # P < 0.05, compared to the previous group.

Article Snippet: A pCMV-based plasmid encoding human CBX3 and a pcDNA3-based plasmid encoding human NFE2L2 were obtained from the nonprofit plasmid repository, Addgene (Cambridge, MA, USA).

Techniques: Sequencing, Expressing, Immunohistochemical staining, Staining, Western Blot, Transfection

Journal: Oncogene

Article Title: CBX3 promotes multidrug resistance by suppressing ferroptosis in colorectal carcinoma via the CUL3/NRF2/GPX2 axis

doi: 10.1038/s41388-025-03337-9

Figure Lengend Snippet: A qRT-PCR assays of NRF2 mRNA expression in SW480-Con/CBX3 cells. B Co-immunoprecipitation assays of NRF2 ubiquitination in CRC cells treated with siCBX3 and Ub-HA plasmids, followed by MG132 incubation. C qRT-PCR and western blotting to detect CUL3 mRNA and protein expression in SW480-Con/CBX3 cells. D ChIP and Luciferase reporter assays of CBX3 binding domain in the promoter of CUL3. HEK-293T cells were transfected with CBX3 plasmid, the cell lysates were incubated with CBX3 antibodies, and qRT-PCR was used to confirm the binding of CBX3 to the CUL3 promoter. Furthermore, firefly luciferase reporter assays were used to analyze the transcriptional activity of CUL3 in SW480-Con/CBX3 cells. E Firefly luciferase reporter assays to analyze the transcriptional activities of truncated CUL3 promoters (chr2:224585276–224585926) in HEK-293T cells with CBX3 overexpression. F Analysis of drug sensitivity in SW480-Con/CBX3 and SW480-CBX3-CUL3 cells after treatment with the indicated drug concentrations. Western blotting was used to detect CBX3 and NRF2 expression in SW480-Con/CBX3/CBX3-CUL3 cells. MTT assays were performed to detect cell viability. Data were presented as the means ± SEM ( N = 5). G GSH/GSSG and MDA assays in SW480-Con/CBX3/CBX3-CUL3 cells following chemotherapy. A – E , G Con: SW480-Con cells, CBX3: SW480-CBX3 cells. CBX3 + CUL3: SW480 stably overexpressed CBX3 and CUL3. Data were presented as the means ± SEM ( N = 3), * P < 0.05, ** P < 0.01, compared with corresponding control groups.

Article Snippet: A pCMV-based plasmid encoding human CBX3 and a pcDNA3-based plasmid encoding human NFE2L2 were obtained from the nonprofit plasmid repository, Addgene (Cambridge, MA, USA).

Techniques: Quantitative RT-PCR, Expressing, Immunoprecipitation, Ubiquitin Proteomics, Incubation, Western Blot, Luciferase, Binding Assay, Transfection, Plasmid Preparation, Activity Assay, Over Expression, Stable Transfection, Control

Journal: Oncogene

Article Title: CBX3 promotes multidrug resistance by suppressing ferroptosis in colorectal carcinoma via the CUL3/NRF2/GPX2 axis

doi: 10.1038/s41388-025-03337-9

Figure Lengend Snippet: A qRT-PCR assays of GPX2 and FTL mRNA expression in SW480-Con/CBX3. B Western blotting to detect the expression of GPX2 and FTL in SW480-Con/CBX3 with and without NRF2 knockdown. C Analysis of GPX2 mRNA expression levels in CRC from the TCGA database. D Western blotting to detect the expression of GPX2 in SW480 cells with and without NRF2 overexpression. E Western blotting to detect the expression of NRF2 and GPX2 in SW480-Con/CBX3 cells with various chemotherapies. F Analysis of drug sensitivity in SW480-Con/CBX3 cells after treatment with the indicated drug concentrations. Western blotting was used to detect the effect of GPX2 knockdown in SW480-Con/CBX3 cells. MTT assays were performed to detect cell viability. G Analysis of GPX2 promoter. The promoter sequences from −2 K to +1 K bp relative to the targeted TSS were scanned using FIMO (reference: https://meme-suite.org/meme/doc/cite.html ) to search for the NFE2L2 motif (MA0150.1), which was obtained from the JASPAR database. H Firefly luciferase reporter assays to analyze the transcriptional activities of GPX2 promoter in SW480-Con/CBX3 cells with and without NRF2 knockdown. A – H Con: SW480-Con cells, CBX3: SW480-CBX3 cells. Data were presented as the means ± SEM ( N = 3). * P < 0.05, ** P < 0.01, compared with corresponding control groups. # P < 0.05, ## P < 0.01, compared with their previous groups.

Article Snippet: A pCMV-based plasmid encoding human CBX3 and a pcDNA3-based plasmid encoding human NFE2L2 were obtained from the nonprofit plasmid repository, Addgene (Cambridge, MA, USA).

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

Journal: Oncogene

Article Title: CBX3 promotes multidrug resistance by suppressing ferroptosis in colorectal carcinoma via the CUL3/NRF2/GPX2 axis

doi: 10.1038/s41388-025-03337-9

Figure Lengend Snippet: A CBX3 overexpression increased Irinotecan resistance in CRC in vivo. Representative images of tumors, tumor size, and weight were obtained 10 days after the initial tumor cell injection ( n = 6). Tumors were measured every two days, and the tumor volumes were calculated. The tumors were excised and weighed 34 days after injection, and representative images of tumors were displayed (bar = 10 mm). B Western blotting to detect the expression of NRF2 and GPX2 in SW480-Con/CBX3 tumor masses with or without Irinotecan treatment. C Immunohistochemical staining of tumor tissues with antibodies against Ki67, NRF2, and GPX2 (low-power view 100×, high-power view 400×, scale bar = 50 μm). D MDA assays in SW480-Con/CBX3 tumor masses with and without chemotherapy. E CBX3 overexpression increases Oxaliplatin resistance in CRC in vivo. Representative images of tumors, tumor size, and weight were obtained at the indicated time point after the initial tumor cell injection ( n = 6). Tumors were measured every two days, and the tumor volumes were calculated. The tumors were excised and weighed 28 days after injection, and representative images of tumors were displayed (bar = 10 mm). A – E Con: SW480-Con cells transplantation tumor masses, CBX3: SW480-CBX3 cells transplantation tumor masses. Data were presented as the means ± SEM, * P < 0.05, ** P < 0.01, compared with corresponding control groups.

Article Snippet: A pCMV-based plasmid encoding human CBX3 and a pcDNA3-based plasmid encoding human NFE2L2 were obtained from the nonprofit plasmid repository, Addgene (Cambridge, MA, USA).

Techniques: Over Expression, In Vivo, Injection, Western Blot, Expressing, Immunohistochemical staining, Staining, Transplantation Assay, Control

Journal: Oncogene

Article Title: CBX3 promotes multidrug resistance by suppressing ferroptosis in colorectal carcinoma via the CUL3/NRF2/GPX2 axis

doi: 10.1038/s41388-025-03337-9

Figure Lengend Snippet: A Immunohistochemistry staining of tumor masses from four PDX samples with CBX3, NRF2, and GPX2 antibodies (low-power view 100×, high-power view 400×, scale bars = 50 μm). B Effects of CBX3/NRF2/GPX2 signaling on irinotecan resistance in the PDX #440 model. Representative images of tumors and tumor sizes were obtained at the indicated time points or 35 days after the initial tumor cell injection ( n = 5). Tumors were measured every two days, and the tumor volumes were calculated. Irinotecan (50 mg/kg/day) was administered from day 45 to day 59. ML385 (30 mg/kg/day) was administered from days 52 to 59. The tumors were excised and weighed 61 days after injection, and representative images of tumors were displayed (bar = 10 mm). The in vivo response to treatment was assessed based on the protocol in “Materials and Methods”. C Immunohistochemical staining of tumor masses after chemotherapy with NRF2, GPX2, and Ki67 antibodies. HE staining of tumor tissues shows different degrees of necrosis and tumor in adjacent tissues (low-power view 100×, high-power view 400×, scale bars = 50 μm). Tumors were excised and weighed 61 days after injection. MDA assays were conducted in tumor masses with or without chemotherapy A – C Con: solvent treatment. Data were presented as the means ± SEM, ** P < 0.01, compared with corresponding control groups. # P < 0.05, compared with Irinotecan-treated groups.

Article Snippet: A pCMV-based plasmid encoding human CBX3 and a pcDNA3-based plasmid encoding human NFE2L2 were obtained from the nonprofit plasmid repository, Addgene (Cambridge, MA, USA).

Techniques: Immunohistochemistry, Staining, Injection, In Vivo, Immunohistochemical staining, Solvent, Control