Journal: bioRxiv

Article Title: Multimodality Molecular Profiling Nominates Targetable Mechanisms in Progressive RV Dysfunction

doi: 10.64898/2026.03.09.710504

Figure Lengend Snippet: RV endothelial and pericyte abundances and colocalization patterns were associated with cardiomyocyte hypoxia signaling and with progressive RV compromise. (A) UMAP visualizing endothelial cells and pericytes in snRNAseq data. (B) Endothelial cell and (C) pericyte relative abundances were similar between mild RVD and control, but elevated in severe RVD. (D) Representative image of RV sections stained with isolectin B4 (IB4) showing capillary density is higher in severe RVD (white arrows: capillaries, yellow: WGA, purple: IB4), quantified as (E) the percent area occupied by endothelial cells. (F) Representative image of RV sections stained with IB4 and platelet derived growth factor receptor beta (PDGFRβ) to mark endothelial cells and pericytes, respectively. Endothelial cell-pericyte localization was altered in severe RVD (white arrows: co-localized endothelial cells and pericytes, red arrow: pericytes not located near endothelial cells, orange: IB4, blue: PDGFRβ) quantified with a colocalization coefficient. (H) Cardiomyocyte HIF1A expression was highest in severe RVD. One way ANOVA with Tukey’s post-hoc, means shown in B-C, E, G.

Article Snippet: Pericyte-endothelial cell colocalization was calculated by staining OCT-embedded RV tissue sections with primary antibody to platelet-derived growth factor receptor beta (PDGFRβ, Cell Signaling Technology, 3169S) at a 1:50 dilution.

Techniques: Control, Staining, Derivative Assay, Expressing

Journal: The American Journal of Pathology

Article Title: Matrix Gla Protein Expression in Pericytes and Myofibroblasts Contributes to Renal Fibrosis

doi: 10.1016/j.ajpath.2025.12.003

Figure Lengend Snippet: Pericytes express Mgp in the kidney. A: Scheme represents Rosa Tomato/+ ;Mgp-Cre mice generation. Cre recombinase expression following the Mgp gene expression deleted the stop cassette and expressed red fluorescent protein (RFP) fluorescence. B: Fluorescence microscopy showing the endogenous RFP expression in tubular interstitial cells of control ( Rosa Tomato/+ ) and Rosa Tomato/+ ;Mgp-Cre kidneys. Nuclei were stained with Hoechst (blue). Bottom panels: Magnified images of yellow dotted boxed areas are presented. C: Mgp target sequence, the sequence of the guide RNA (blue), and single-stranded oligodeoxynucleotide (ssODN) to introduce the desired insertion. The ssODN carries homology arms flanking each side of the target nucleotide with hemagglutinin (HA) sequences ( yellow boxed area ). As shown, the guide RNA anneals on the stop codon (TAG) of Mgp ( red boxed area ), complementary to the protospacer adjacent motif ( green circle ), where co-injected clustered regularly interspaced short palindromic repeats (CRISPR)-associated protein 9 (Cas9) is expected to generate a double-stranded break. D: Left: A representative agarose gel image of genotyping PCR using HA/F3 and HA/R3 primers. Wild-type (WT) Mgp allele produces a 297-bp fragment, and HA- Mgp allele produces a 324-bp fragment. Right: Sanger sequencing analyses confirm the insertion of the HA sequence (yellow) before the stop codon (red). E: Confocal microscopy showing matrix Gla protein (MGP)–expressing cells in a healthy kidney. Tissues were stained using rabbit anti-CD31 or rabbit anti–platelet-derived growth factor receptor (PDGFR)-β with mouse anti-HA antibody, followed by anti-rabbit Alexa 555 (green or white) and anti-mouse Alexa 647 (red) antibodies. Nuclei were stained with Hoechst (blue). Right: Magnified images of yellow dotted boxed areas are presented. Scale bars: 10 μm ( B , bottom panels, and E , left ); 20 μm ( B , top panels ); 2 μm ( E , right ). IRES, internal ribosome entry site.

Article Snippet: The following primary antibodies were used: anti-HA tag antibody (1:50; 2367; Cell Signaling Technology, Danvers, MA), anti-CD31 (1:200; 77699; Cell Signaling Technology), anti–platelet-derived growth factor receptor β (1:50; 3169; Cell Signaling Technology), and anti–α smooth muscle actin (SMA) (1:200; 19245; Cell Signaling Technology).

Techniques: Expressing, Gene Expression, Fluorescence, Microscopy, Control, Staining, Sequencing, Introduce, Injection, CRISPR, Agarose Gel Electrophoresis, Confocal Microscopy, Derivative Assay

Journal: The American Journal of Pathology

Article Title: Matrix Gla Protein Expression in Pericytes and Myofibroblasts Contributes to Renal Fibrosis

doi: 10.1016/j.ajpath.2025.12.003

Figure Lengend Snippet: Loss of matrix Gla protein (MGP) suppresses the extracellular matrix expression and kidney fibrosis. A: Sirius red–stained histologic sections and quantification of stained area confirmed the collagen fibers in wild-type (WT) or Mgp –/– kidneys at 2 weeks after the folic acid (FA) administration. For quantification, 10 fields were examined from each sample. B: Quantitative real-time PCR analysis of fibrotic marker gene expression levels in WT and Mgp –/– kidneys before and after FA (or vehicle) administration in reference to Hprt . C: Confocal microscopy showing pericytes ( arrowheads ) in the WT and Mgp –/– kidneys. Tissues were stained using rabbit anti–platelet-derived growth factor receptor (PDGFR)-β, followed by anti-rabbit Alexa 555 (red). Nuclei were stained with Hoechst (blue). Right: The bar graph represents the number of pericytes over total interstitial cells per field. For each sample, 10 fields were examined. D: Quantitative real-time PCR analysis of Hey1 expression levels in WT and Mgp –/– kidneys before or 2 weeks after the FA administration in reference to Hprt . A – D: Statistical analysis: t -test ( A and C ) or two-way analysis of variance, followed by Bonferroni correction ( B and D ), was used to calculate P . The results are presented as means ± SEM ( A – D ). n = 4 for each group ( A and C ); n = 3 for each group ( B ); n = 3 to 5 for each group ( D ). ∗ P < 0.05, ∗∗ P < 0.01, and ∗∗∗∗ P < 0.0001. Scale bars: 100 μm ( A , left ); 20 μm ( C ). HPF, high-power field; NS, not significant.

Article Snippet: The following primary antibodies were used: anti-HA tag antibody (1:50; 2367; Cell Signaling Technology, Danvers, MA), anti-CD31 (1:200; 77699; Cell Signaling Technology), anti–platelet-derived growth factor receptor β (1:50; 3169; Cell Signaling Technology), and anti–α smooth muscle actin (SMA) (1:200; 19245; Cell Signaling Technology).

Techniques: Expressing, Staining, Real-time Polymerase Chain Reaction, Marker, Gene Expression, Confocal Microscopy, Derivative Assay

Journal: Rsc Applied Polymers

Article Title: Biodegradable copper complexing polymeric microparticles relieve oxidative stress

doi: 10.1039/d5lp00289c

Figure Lengend Snippet: In vitro assessment of PASmp and Cu-PASmp on HCF cells. (A) Viability (%) of HCF cells upon 24 hour incubation with PASmp, Cu-PASmp or Cu(OAc) 2 determined by the MTT assay. (B) Oxidative stress rescue capacity of 300 μg mL −1 PASmp, Cu-PASmp, or 60 μM Cu(OAc) 2 upon treatment of HCF with 10 µM H 2 O 2 for 24 h assessed via the MTT assay. (C) Oxidative stress rescue of HCF fluorescence microscopy stained with DAPI (blue) and phalloidin (red). All samples are incubated with 10 µM H 2 O 2 . Scale bars are 300 μm. (D) SOD1 expression levels upon oxidative stress rescue conditions determined by western blotting, gel image (1) and quantification plot (2). All quantitative western data acquired at the same exposure time. Data in (A), (B), and (D2) represent mean ± SD ( n = 3). Significant difference was determined by Welch's t -test: * for p < 0.05, ** for p < 0.01, and *** for p < 0.001.

Article Snippet: The expression level of SOD1 protein is quantified by enzyme associated fluorescence after tagging with rabbit-derived SOD1 antibody (1 : 1000, Cell Signaling Technology, MA, USA, Cat. no. 2770).

Techniques: In Vitro, Incubation, MTT Assay, Fluorescence, Microscopy, Staining, Expressing, 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 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