polyclonal rabbit derived zc3h15 antibody  (Proteintech)

Journal: Cell Death & Disease

Article Title: ZC3H15 regulates the ubiquitination of PTEN via recruitment of TRIM56 and promotes malignant progression of non-small cell lung cancer

doi: 10.1038/s41419-025-08138-2

Figure Lengend Snippet: A Venn diagrams showing overlaps between five GEO datasets and the data from TCGA database. B ZC3H15 mRNA expression was investigated in the TCGA database and compared among 33 kinds of tumors. C The analysis of overall survival in all NSCLC patients. D Western blotting analyses of ZC3H15 levels in eight lung cancer tissues and matched normal tissues. Student’s t test. Mean ± SD, n = 3. ** P < 0.01. E ZC3H15 levels in normal alveolar(Ⅰ) and bronchial epithelial cell(Ⅱ), adenocarcinoma(Ⅲ),and squamous cell carcinoma(Ⅳ) using immunohistochemistry. Magnification: ×200.

Article Snippet: Slides were incubated overnight with polyclonal rabbit-derived ZC3H15 antibody (26241-1-AP, 1:200, Proteintech, China) at 4 °C.

Techniques: Expressing, Western Blot, Immunohistochemistry

Journal: Cell Death & Disease

Article Title: ZC3H15 regulates the ubiquitination of PTEN via recruitment of TRIM56 and promotes malignant progression of non-small cell lung cancer

doi: 10.1038/s41419-025-08138-2

Figure Lengend Snippet: A Immunofluorescence was performed to detect ZC3H15 localization in cell lines. B Western blotting analyses of ZC3H15 expression in lung epithelial cell line, HBE and six NSCLC cell lines. Student’s t test. Mean ± SD, n = 3. *** P < 0.001. C Western blotting analyzing the expression of ZC3H15 in the indicated cells. Multiple t tests. Mean ± SD, n = 3. *** P < 0.001. D Cell viability was analyzed by CCK8. Multiple t tests. Mean ± SD, n = 3. ** P < 0.01. E Cell growth was determined by colony formation. Mean ± SD, n = 3. ** P < 0.01, *** P < 0.001. F DNA replication of A549 cells was determined by EDU staining. Scale bar: 20 μm. Mean ± SD, n = 3. *** P < 0.001. G The xenograft tumor model in nude mice established by A549 cells with ZC3H15 overexpression. Student’s t test. Mean ± SD, n = 3. *** P < 0.001. Cell migration ( H ) and invasion ( I ) was evaluated by the Transwell migration assay; cells that migrated to the lower chamber were stained with hematoxylin and counted. Mean ± SD, n = 3. *** P < 0.001.

Article Snippet: Slides were incubated overnight with polyclonal rabbit-derived ZC3H15 antibody (26241-1-AP, 1:200, Proteintech, China) at 4 °C.

Techniques: Immunofluorescence, Western Blot, Expressing, Staining, Over Expression, Migration, Transwell Migration Assay

Journal: Cell Death & Disease

Article Title: ZC3H15 regulates the ubiquitination of PTEN via recruitment of TRIM56 and promotes malignant progression of non-small cell lung cancer

doi: 10.1038/s41419-025-08138-2

Figure Lengend Snippet: A GSEA in the TCGA database of ZC3H15-related enrichment plots were performed. B Western blotting analyzing the expression of proteins involved in the AKT-mTOR pathway in A549 and H1299 cells treated with DMSO or the AKT pathway inhibitor LY294002. C Cell growth was determined by colony formation. Unpaired t test. Mean ± SD, n = 3. *** P < 0.001. D DNA replication of A549 and H1299 cells treated with DMSO or the AKT pathway inhibitor LY294002 was determined by EDU staining. Scale bar: 200 μm. Mean ± SD, n = 3. *** P < 0.001. E Cell migration was evaluated by the Transwell migration assay. Mean ± SD, n = 3. ** P < 0.01, *** P < 0.001. F Wound-healing assay showing the LY294002 effect on the ZC3H15 overexpressed A549 and H1299 cell migration. Mean ± SD, n = 3. ** P < 0.01, *** P < 0.001.

Article Snippet: Slides were incubated overnight with polyclonal rabbit-derived ZC3H15 antibody (26241-1-AP, 1:200, Proteintech, China) at 4 °C.

Techniques: Western Blot, Expressing, Staining, Migration, Transwell Migration Assay, Wound Healing Assay

Journal: Cell Death & Disease

Article Title: ZC3H15 regulates the ubiquitination of PTEN via recruitment of TRIM56 and promotes malignant progression of non-small cell lung cancer

doi: 10.1038/s41419-025-08138-2

Figure Lengend Snippet: A The Venn diagram shows the overlaps between protein mass spectrometry analysis results and the data from BioGRID website. B The raw MS/MS spectrum of PTEN. C Immunofluorescence assay results indicate that ZC3H15 and PTEN colocalize within the cytoplasm of A549 and H1299 cells. D , E Interactions between PHF23 and ACTN4 in A549 and H1299 cells measured by co-immunoprecipitation. F Cell growth of A549 treated with DMSO or VO-Ohpic was determined by colony formation. Unpaired t test. Mean ± SD, n = 3. *** P < 0.001. G DNA replication of A549 treated with DMSO or VO-Ohpic was determined by EDU staining. Scale bar: 20 μm. Mean ± SD, n = 3. ** P < 0.01, *** P < 0.001. H Cell migration and invasion of A549 was evaluated by the Transwell migration assay. Mean ± SD, n = 3. ** P < 0.01, *** P < 0.001. I The migration of A549 treated with DMSO or VO-Ohpic was evaluated by the wound-healing assay. Mean ± SD, n = 3. *** P < 0.001.

Article Snippet: Slides were incubated overnight with polyclonal rabbit-derived ZC3H15 antibody (26241-1-AP, 1:200, Proteintech, China) at 4 °C.

Techniques: Mass Spectrometry, Tandem Mass Spectroscopy, Immunofluorescence, Immunoprecipitation, Staining, Migration, Transwell Migration Assay, Wound Healing Assay

Journal: Cell Death & Disease

Article Title: ZC3H15 regulates the ubiquitination of PTEN via recruitment of TRIM56 and promotes malignant progression of non-small cell lung cancer

doi: 10.1038/s41419-025-08138-2

Figure Lengend Snippet: A Schematic diagram of ZC3H15 splicing mutants. B The expression of myc-tag after transfected with ZC3H15 splicing mutant cDNA. C Interactions between ZC3H15 splicing mutants and PTEN in A549 and H1299 cells measured by co-immunoprecipitation. D Cell viability of A549 and H1299 transfected with ZC3H15 cDNA or ZC3H15 MUT3 cDNA was analyzed by CCK8. Unpaired t test. Mean ± SD, n = 3. *** P < 0.001. E Cell growth of A549 transfected with ZC3H15 cDNA or ZC3H15 MUT3 cDNA was determined by colony formation. Mean ± SD, n = 3. *** P < 0.001. F DNA replication of A549 transfected with ZC3H15 cDNA or ZC3H15 MUT3 cDNA was determined by EDU staining. Scale bar: 20 μm. Mean ± SD, n = 3. *** P < 0.001. The migration ( G ) and invasion ( H ) of A549 transfected with ZC3H15 cDNA or ZC3H15 MUT3 cDNA was evaluated by the Transwell migration assay. Mean ± SD, n = 3. *** P < 0.001. I Western blotting analyzing the expression of proteins involved in the AKT-mTOR pathway and proliferation- and migration-related proteins in A549 and H1299 cells transfected with ZC3H15 cDNA or ZC3H15 MUT3 cDNA.

Article Snippet: Slides were incubated overnight with polyclonal rabbit-derived ZC3H15 antibody (26241-1-AP, 1:200, Proteintech, China) at 4 °C.

Techniques: Expressing, Transfection, Mutagenesis, Immunoprecipitation, Staining, Migration, Transwell Migration Assay, Western Blot

Journal: Cell Death & Disease

Article Title: ZC3H15 regulates the ubiquitination of PTEN via recruitment of TRIM56 and promotes malignant progression of non-small cell lung cancer

doi: 10.1038/s41419-025-08138-2

Figure Lengend Snippet: A Levels of PTEN ubiquitination were evaluated by immunoprecipitation using an anti-PTEN antibody, followed by anti-HA immunoblotting. B , C Levels of PTEN ubiquitination of A549 and H1299 transfected with K48/K63 mutant Ub were evaluated by immunoprecipitation using an anti-PTEN antibody, followed by anti-HA immunoblotting. D 3D Binding model analysis (ZC3H15 in pink, TRIM56 in blue and PTEN in cyan). The key residues are shown as sticks. H-bonds are shown as yellow dashed lines. E Confocal microscopy of triplestained Flag-ZC3H15(green), TRIM56(red) and DAPI(blue) in H1299 cells. F Confocal microscopy of triplestained GFP-PTEN(green), TRIM56(red) and DAPI(blue) in H1299 cells. G Interactions between TRIM56 with Flag-ZC3H15 and GFP-PTEN in A549 and H1299 cells measured by co-immunoprecipitation. H Levels of PTEN ubiquitination of A549 and H1299 with TRIM56 knockdown were evaluated by immunoprecipitation using an anti-PTEN antibody, followed by anti-HA immunoblotting. I Levels of PTEN ubiquitination were evaluated by immunoprecipitation using an anti-PTEN antibody, followed by anti-HA immunoblotting.

Article Snippet: Slides were incubated overnight with polyclonal rabbit-derived ZC3H15 antibody (26241-1-AP, 1:200, Proteintech, China) at 4 °C.

Techniques: Ubiquitin Proteomics, Immunoprecipitation, Western Blot, Transfection, Mutagenesis, Binding Assay, Confocal Microscopy, Knockdown

Journal: Cell Death & Disease

Article Title: ZC3H15 regulates the ubiquitination of PTEN via recruitment of TRIM56 and promotes malignant progression of non-small cell lung cancer

doi: 10.1038/s41419-025-08138-2

Figure Lengend Snippet: A IC50 analysis of NSCLC patients with high and low ZC3H15 expression based on TCGA data. B Western blotting analyzing the expression of PTEN and p-PTEN in A549 and A549-DDP. C Viability of A549 and A549-DDP cells was analyzed by CCK8 24 h after treatment with different concentrations of cisplatin. D Viability of A549 and H1299 cells with ZC3H15 overexpressed was analyzed by CCK8 24 h after treatment with different concentrations of cisplatin. E Representative explanted tumor growth curve of mice treated as indicated ( n = 5 per group). F Xenograft tumors from H1299 cells. G Tumor growth curves were shown. Mean ± SD, n = 5. *** P < 0.001. H Quantification of xenograft tumor weights. Mean ± SD, n = 5. *** P < 0.001. I Representative pictures of H&E and IHC staining of ZC3H15, Ki-67, and PTEN in the indicated xenograft tumors. Scale bar, 200 μm.

Article Snippet: Slides were incubated overnight with polyclonal rabbit-derived ZC3H15 antibody (26241-1-AP, 1:200, Proteintech, China) at 4 °C.

Techniques: Expressing, Western Blot, Immunohistochemistry

Journal: Cell Death & Disease

Article Title: ZC3H15 regulates the ubiquitination of PTEN via recruitment of TRIM56 and promotes malignant progression of non-small cell lung cancer

doi: 10.1038/s41419-025-08138-2

Figure Lengend Snippet: ZC3H15 promotes the proliferation, migration, invasion and chemotherapy resistance of NSCLC by mediating PTEN ubiquitination degradation and activating the AKT-mTOR signaling pathway.

Article Snippet: Slides were incubated overnight with polyclonal rabbit-derived ZC3H15 antibody (26241-1-AP, 1:200, Proteintech, China) at 4 °C.

Techniques: Migration, Ubiquitin Proteomics