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cell staining reagents  (Beyotime)


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    Structured Review

    Beyotime cell staining reagents
    Cell Staining Reagents, supplied by Beyotime, used in various techniques. Bioz Stars score: 99/100, based on 33153 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/cells+with+dapi/DAPI/pmc13098730-37-0-14
    Average 99 stars, based on 33153 article reviews
    cell staining reagents - by Bioz Stars, 2026-11
    99/100 stars

    Images

    Related Articles

    Staining:

    Article Title: SATB1 lowers the T cell receptor signaling threshold for thymocyte positive selection via super-enhancer reprogramming.
    Article Snippet: Followed by 3 × PBS wash, followed by incubation with a secondary antibody in conjunctionwithAlexa 647fluorescence (FD0144) 1:1000 in 1 × PBS for onehour in the dark. .. Afterwashing three times, the nuclei were stained with 10 μg/ml DAPI (Beyotime). .. Images were acquired by a confocal microscope (Carl Zeiss, LSM 880 with Airyscan or Nikon A1 R) with a 100x objective.

    Article Title: CD147 drives keloid formation by promoting fibroblast proliferation via the CENPF signaling pathway.
    Article Snippet: After being inoculated onto 24-well culture plates and subjected to the indicated treatments, the CoraLite488 TUNEL assay apoptosis detection kit (Proteintech, Wuhan, China) was used to detect apoptotic cells according to the manufacturer’s instructions. .. Nuclei were stained with 4′,6- diamidino-2-phenylindole (DAPI, Beyotime, Shanghai, China) for 5 min. TUNEL-positive apoptotic cells were labeled with green fluorescence and visualized by a Leica Microsystems microscope (Solms, Germany). ..

    Article Title: The AGEs/RAGE Axis Contributes to Myocardial Cell Death in Type 1 Diabetic Cardiomyopathy via the ROS/NF‐ κ B/NLRP3 Inflammasome Pathway
    Article Snippet: β ‐Actin (Cat#R1207‐1) was purchased from Huabio (Harbin, China). .. Trypsin solution (Cat#C0201), CM‐H2DCFDA staining (Cat#S0035S), Cy3‐conjugated secondary antibody (Cat# A0521), FITC‐conjugated secondary antibody (Cat# A0568), 4 ′ ,6‐diamidino‐2‐phenylindole (DAPI) (CatC1005), RIPA (Cat#P0013B), protein SDS‐PAGE protein loading buffer (5X) (Cat#P0015), BCA protein concentration measurement kit (Cat#P0011), and lactate dehydrogenase (LDH) cytotoxicity assay kit (Cat#C0016) were purchased from Beyotime (Beijing, China). .. α ‐Actinin (Cat#GTX11007) was from GeneTex (Texas, United States).

    Blocking Assay:

    Article Title: NSUN3‐Associated m5C Modification of tRF5‐GlyTCC Suppresses Colorectal Cancer Progression via RPL23‐Linked Nucleolar Stress and p53‐Associated Autophagy and Enhances 5‐FU Sensitivity
    Article Snippet: .. Cells were fixed with 4% paraformaldehyde for 15 min and permeabilized with 0.1% Triton X‐100 for 10 min. After blocking with 5% BSA for 1 h, cells were incubated with an anti‐NPM1 antibody (Abcam, UK) overnight at 4°C, followed by incubation with an Alexa Fluor‐conjugated secondary antibody for 1 h. Nuclei were counterstained with DAPI (Beyotime, China). ..

    Incubation:

    Article Title: NSUN3‐Associated m5C Modification of tRF5‐GlyTCC Suppresses Colorectal Cancer Progression via RPL23‐Linked Nucleolar Stress and p53‐Associated Autophagy and Enhances 5‐FU Sensitivity
    Article Snippet: .. Cells were fixed with 4% paraformaldehyde for 15 min and permeabilized with 0.1% Triton X‐100 for 10 min. After blocking with 5% BSA for 1 h, cells were incubated with an anti‐NPM1 antibody (Abcam, UK) overnight at 4°C, followed by incubation with an Alexa Fluor‐conjugated secondary antibody for 1 h. Nuclei were counterstained with DAPI (Beyotime, China). ..

    Article Title: Lineage-specific and gestational age–dependent DNA methylation programming orchestrates human placental development
    Article Snippet: Then, they were incubated with 5mC (61255, Active Motif), EGFR (ab30, Abcam), HLA-G (ab52454, Abcam), TFAP2C (sc12762, Santa Cruz Biotechnology), CDH2 (ab18203, Abcam), EpCAM (MA5-12436, Thermo Fisher Scientific), TP63 (ab124762, Abcam), SDC1 (67155-1-Ig, Proteintech), CGB (ab9582, Abcam), or α-SMA (MA1-06110, Thermo Fisher Scientific) overnight at 4°C. .. After multiple washes to remove residual primary antibody, cells or sections were then incubated with AF488/AF594-conjugated secondary antibodies (Thermo Fisher Scientific) for 1 hour at room temperature and mounted in prolonged anti-fade reagent with 4′,6-diamidino-2-phenylindole (DAPI, Beyotime). .. Confocal imaging was performed with Zeiss LSM 880 confocal microscopes or LEICA TCS SP8, and analyzed with Zeiss Zen blue edition.

    TUNEL Assay:

    Article Title: CD147 drives keloid formation by promoting fibroblast proliferation via the CENPF signaling pathway.
    Article Snippet: After being inoculated onto 24-well culture plates and subjected to the indicated treatments, the CoraLite488 TUNEL assay apoptosis detection kit (Proteintech, Wuhan, China) was used to detect apoptotic cells according to the manufacturer’s instructions. .. Nuclei were stained with 4′,6- diamidino-2-phenylindole (DAPI, Beyotime, Shanghai, China) for 5 min. TUNEL-positive apoptotic cells were labeled with green fluorescence and visualized by a Leica Microsystems microscope (Solms, Germany). ..

    Labeling:

    Article Title: CD147 drives keloid formation by promoting fibroblast proliferation via the CENPF signaling pathway.
    Article Snippet: After being inoculated onto 24-well culture plates and subjected to the indicated treatments, the CoraLite488 TUNEL assay apoptosis detection kit (Proteintech, Wuhan, China) was used to detect apoptotic cells according to the manufacturer’s instructions. .. Nuclei were stained with 4′,6- diamidino-2-phenylindole (DAPI, Beyotime, Shanghai, China) for 5 min. TUNEL-positive apoptotic cells were labeled with green fluorescence and visualized by a Leica Microsystems microscope (Solms, Germany). ..

    Fluorescence:

    Article Title: CD147 drives keloid formation by promoting fibroblast proliferation via the CENPF signaling pathway.
    Article Snippet: After being inoculated onto 24-well culture plates and subjected to the indicated treatments, the CoraLite488 TUNEL assay apoptosis detection kit (Proteintech, Wuhan, China) was used to detect apoptotic cells according to the manufacturer’s instructions. .. Nuclei were stained with 4′,6- diamidino-2-phenylindole (DAPI, Beyotime, Shanghai, China) for 5 min. TUNEL-positive apoptotic cells were labeled with green fluorescence and visualized by a Leica Microsystems microscope (Solms, Germany). ..

    Microscopy:

    Article Title: CD147 drives keloid formation by promoting fibroblast proliferation via the CENPF signaling pathway.
    Article Snippet: After being inoculated onto 24-well culture plates and subjected to the indicated treatments, the CoraLite488 TUNEL assay apoptosis detection kit (Proteintech, Wuhan, China) was used to detect apoptotic cells according to the manufacturer’s instructions. .. Nuclei were stained with 4′,6- diamidino-2-phenylindole (DAPI, Beyotime, Shanghai, China) for 5 min. TUNEL-positive apoptotic cells were labeled with green fluorescence and visualized by a Leica Microsystems microscope (Solms, Germany). ..

    BIA-KA:

    Article Title: The AGEs/RAGE Axis Contributes to Myocardial Cell Death in Type 1 Diabetic Cardiomyopathy via the ROS/NF‐ κ B/NLRP3 Inflammasome Pathway
    Article Snippet: β ‐Actin (Cat#R1207‐1) was purchased from Huabio (Harbin, China). .. Trypsin solution (Cat#C0201), CM‐H2DCFDA staining (Cat#S0035S), Cy3‐conjugated secondary antibody (Cat# A0521), FITC‐conjugated secondary antibody (Cat# A0568), 4 ′ ,6‐diamidino‐2‐phenylindole (DAPI) (CatC1005), RIPA (Cat#P0013B), protein SDS‐PAGE protein loading buffer (5X) (Cat#P0015), BCA protein concentration measurement kit (Cat#P0011), and lactate dehydrogenase (LDH) cytotoxicity assay kit (Cat#C0016) were purchased from Beyotime (Beijing, China). .. α ‐Actinin (Cat#GTX11007) was from GeneTex (Texas, United States).

    Protein Concentration:

    Article Title: The AGEs/RAGE Axis Contributes to Myocardial Cell Death in Type 1 Diabetic Cardiomyopathy via the ROS/NF‐ κ B/NLRP3 Inflammasome Pathway
    Article Snippet: β ‐Actin (Cat#R1207‐1) was purchased from Huabio (Harbin, China). .. Trypsin solution (Cat#C0201), CM‐H2DCFDA staining (Cat#S0035S), Cy3‐conjugated secondary antibody (Cat# A0521), FITC‐conjugated secondary antibody (Cat# A0568), 4 ′ ,6‐diamidino‐2‐phenylindole (DAPI) (CatC1005), RIPA (Cat#P0013B), protein SDS‐PAGE protein loading buffer (5X) (Cat#P0015), BCA protein concentration measurement kit (Cat#P0011), and lactate dehydrogenase (LDH) cytotoxicity assay kit (Cat#C0016) were purchased from Beyotime (Beijing, China). .. α ‐Actinin (Cat#GTX11007) was from GeneTex (Texas, United States).

    LDH Cytotoxicity Assay:

    Article Title: The AGEs/RAGE Axis Contributes to Myocardial Cell Death in Type 1 Diabetic Cardiomyopathy via the ROS/NF‐ κ B/NLRP3 Inflammasome Pathway
    Article Snippet: β ‐Actin (Cat#R1207‐1) was purchased from Huabio (Harbin, China). .. Trypsin solution (Cat#C0201), CM‐H2DCFDA staining (Cat#S0035S), Cy3‐conjugated secondary antibody (Cat# A0521), FITC‐conjugated secondary antibody (Cat# A0568), 4 ′ ,6‐diamidino‐2‐phenylindole (DAPI) (CatC1005), RIPA (Cat#P0013B), protein SDS‐PAGE protein loading buffer (5X) (Cat#P0015), BCA protein concentration measurement kit (Cat#P0011), and lactate dehydrogenase (LDH) cytotoxicity assay kit (Cat#C0016) were purchased from Beyotime (Beijing, China). .. α ‐Actinin (Cat#GTX11007) was from GeneTex (Texas, United States).



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    Image Search Results


    ( a ) Frequency of CD8 + T cells among live lymphocytes in human lung, brain frontal lobe and leptomeninges. ( b ) CD69 and CD103 co-expression among CD8 + T cells across tissues (n=5; bars represent mean ± SEM). ( c ) Flow cytometry of combinatorial dual tetramer staining with Epstein Barr virus (EBV, left) and influenza A virus (IAV, right) HLA-A*02 tetramers from two donors (rows). Gated on live CD3 + CD8 + CD4 - T cells negative for background single fluorophore staining and irrelevant tetramer fluorophores. ( d ) Schematic of post-mortem (n=5) and surgical resection (n=3) cohorts, experimental workflow, and hierarchy of antigen-specific T cell identification methods stratified by confidence level. ( e ) Number of virus-specific CD8+ T cells recovered from the leptomeninges and brain tissue for each donor, stratified by identification method and donor cohort where SR=surgical resection (top) and PM=postmortem (bottom). Brain tissue for the PM cohort encompasses frontal lobe, hippocampus, and cerebellum. Dots represent the absolute number of T cells per specificity, and stacked bars are the cumulative total per donor. All tissues were stained with HLA-A*02 tetramers to EBV, cytomegalovirus (CMV), IAV, and SARS-CoV-2 (COVID) with the exception of PM001 (§) which was stained for EBV tetramer only. ND= none detected. Full tissue-level counts provided in Supplementary Table 2.

    Journal: bioRxiv

    Article Title: Common viral infections seed regionally distinct resident memory T cells in the human CNS

    doi: 10.64898/2026.09.21.753190

    Figure Lengend Snippet: ( a ) Frequency of CD8 + T cells among live lymphocytes in human lung, brain frontal lobe and leptomeninges. ( b ) CD69 and CD103 co-expression among CD8 + T cells across tissues (n=5; bars represent mean ± SEM). ( c ) Flow cytometry of combinatorial dual tetramer staining with Epstein Barr virus (EBV, left) and influenza A virus (IAV, right) HLA-A*02 tetramers from two donors (rows). Gated on live CD3 + CD8 + CD4 - T cells negative for background single fluorophore staining and irrelevant tetramer fluorophores. ( d ) Schematic of post-mortem (n=5) and surgical resection (n=3) cohorts, experimental workflow, and hierarchy of antigen-specific T cell identification methods stratified by confidence level. ( e ) Number of virus-specific CD8+ T cells recovered from the leptomeninges and brain tissue for each donor, stratified by identification method and donor cohort where SR=surgical resection (top) and PM=postmortem (bottom). Brain tissue for the PM cohort encompasses frontal lobe, hippocampus, and cerebellum. Dots represent the absolute number of T cells per specificity, and stacked bars are the cumulative total per donor. All tissues were stained with HLA-A*02 tetramers to EBV, cytomegalovirus (CMV), IAV, and SARS-CoV-2 (COVID) with the exception of PM001 (§) which was stained for EBV tetramer only. ND= none detected. Full tissue-level counts provided in Supplementary Table 2.

    Article Snippet: Viable (DAPI-negative) CD8 + CD4 - cells were sorted to 80% purity on a Sony SH800S cell sorter.

    Techniques: Expressing, Flow Cytometry, Staining, Virus, Tissue, Ex Vivo, FACS, Single Cell

    CircATXN2 is upregulated in PAH models and hypoxic PASMCs A Right ventricular systolic pressure (RVSP) measured by invasive catheterization in control (Ctrl, normoxia) and Su-Hyp-induced PAH (PAH) mice ( n = 6). B , C Volcano plot (B) and Heatmap (C) of differentially expressed circRNAs from RNA-seq analysis of lung tissue from Ctrl and PAH mice (fold change (> 2 or < 0.5), P < 0.05). D qRT-PCR analysis of circATXN2 expression in primary mouse pulmonary artery endothelial cells (PAECs) versus pulmonary artery smooth muscle cells (PASMCs) ( n = 3). E qRT-PCR analysis of circATXN2 expression in PASMCs exposed to normoxia (Nor) or hypoxia (Hyp, 3% O2 for 24 h) ( n = 3) F RT-PCR validation of circATXN2 in PASMCs using divergent primers (amplifying circATXN2) and convergent primers (amplifying linear ATXN2 mRNA) with cDNA and genomic DNA (gDNA) templates. 18S was used as a positive control for both templates. G qRT-PCR analysis of circATXN2 and linear ATXN2 mRNA levels in PASMCs treated with RNase R or a mock control ( n = 3). H Schematic and Sanger sequencing validation of the back-splicing junction (BSJ) of circATXN2, confirming its origin from exons 18 and 19. I Fluorescence in situ hybridization (FISH) of circATXN2 (red) and 18S (green) in PASMCs, demonstrating primary cytoplasmic localization. Nuclei were counterstained with DAPI (blue). Scale bar: 50 µm. J RT-PCR analysis of the distribution of circATXN2 in diverse organs. K Conservation analysis of the circATXN2 sequence in different species. All data are presented as the mean ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001 , ****P < 0.0001, ns , not significant

    Journal: Respiratory Research

    Article Title: CircATXN2 exacerbates pulmonary arterial hypertension by modulating the miR-138-5p/SMURF1 axis

    doi: 10.1186/s12931-026-03647-w

    Figure Lengend Snippet: CircATXN2 is upregulated in PAH models and hypoxic PASMCs A Right ventricular systolic pressure (RVSP) measured by invasive catheterization in control (Ctrl, normoxia) and Su-Hyp-induced PAH (PAH) mice ( n = 6). B , C Volcano plot (B) and Heatmap (C) of differentially expressed circRNAs from RNA-seq analysis of lung tissue from Ctrl and PAH mice (fold change (> 2 or < 0.5), P < 0.05). D qRT-PCR analysis of circATXN2 expression in primary mouse pulmonary artery endothelial cells (PAECs) versus pulmonary artery smooth muscle cells (PASMCs) ( n = 3). E qRT-PCR analysis of circATXN2 expression in PASMCs exposed to normoxia (Nor) or hypoxia (Hyp, 3% O2 for 24 h) ( n = 3) F RT-PCR validation of circATXN2 in PASMCs using divergent primers (amplifying circATXN2) and convergent primers (amplifying linear ATXN2 mRNA) with cDNA and genomic DNA (gDNA) templates. 18S was used as a positive control for both templates. G qRT-PCR analysis of circATXN2 and linear ATXN2 mRNA levels in PASMCs treated with RNase R or a mock control ( n = 3). H Schematic and Sanger sequencing validation of the back-splicing junction (BSJ) of circATXN2, confirming its origin from exons 18 and 19. I Fluorescence in situ hybridization (FISH) of circATXN2 (red) and 18S (green) in PASMCs, demonstrating primary cytoplasmic localization. Nuclei were counterstained with DAPI (blue). Scale bar: 50 µm. J RT-PCR analysis of the distribution of circATXN2 in diverse organs. K Conservation analysis of the circATXN2 sequence in different species. All data are presented as the mean ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001 , ****P < 0.0001, ns , not significant

    Article Snippet: After washing with PBS, the sections were incubated with a Cy3-conjugated anti-rabbit at RT for 1 h, followed by labeling the cell nuclei with DAPI (Servicebio).

    Techniques: Control, RNA Sequencing, Quantitative RT-PCR, Expressing, Reverse Transcription Polymerase Chain Reaction, Biomarker Discovery, Positive Control, Sequencing, Fluorescence, In Situ Hybridization

    CircATXN2 knockdown alleviates hypoxia-induced PASMC proliferation and migration. PASMCs were transfected with a negative control siRNA (si-NC) or an siRNA targeting circATXN2 (si-circATXN2) for 24 h. Cells were then exposed to normoxia (Nor) or hypoxia (Hyp, 3% O2) for an additional 24 h. A Viability measured by Cell Count Kit-8 (CCK-8) assays with transfected with si-circATXN2 ( n = 5) B , E Representative images ( B ) and quantification ( E ) of cell proliferation assessed by EdU incorporation. EdU-positive (proliferating) cells are green; nuclei are counterstained with DAPI (blue) ( n = 6). Scale bar: 100 µm. C , F Representative images ( C ) and quantification ( F ) of cell migration assessed by a wound healing assay. Images were captured at 0 h and 24 h post-scratch ( n = 5). Scale bar: 100 µm. D , G Representative images ( D ) and quantification ( G ) of cell migration assessed by a Transwell assay. Migrated cells were stained with crystal violet ( n = 5). Scale bar: 50 µm. All data are presented as the mean ± SEM. Statistical significance was determined by one-way ANOVA followed by Tukey's post-hoc test. * P < 0.05, **P < 0.01, ***P < 0.001, ns , not significant

    Journal: Respiratory Research

    Article Title: CircATXN2 exacerbates pulmonary arterial hypertension by modulating the miR-138-5p/SMURF1 axis

    doi: 10.1186/s12931-026-03647-w

    Figure Lengend Snippet: CircATXN2 knockdown alleviates hypoxia-induced PASMC proliferation and migration. PASMCs were transfected with a negative control siRNA (si-NC) or an siRNA targeting circATXN2 (si-circATXN2) for 24 h. Cells were then exposed to normoxia (Nor) or hypoxia (Hyp, 3% O2) for an additional 24 h. A Viability measured by Cell Count Kit-8 (CCK-8) assays with transfected with si-circATXN2 ( n = 5) B , E Representative images ( B ) and quantification ( E ) of cell proliferation assessed by EdU incorporation. EdU-positive (proliferating) cells are green; nuclei are counterstained with DAPI (blue) ( n = 6). Scale bar: 100 µm. C , F Representative images ( C ) and quantification ( F ) of cell migration assessed by a wound healing assay. Images were captured at 0 h and 24 h post-scratch ( n = 5). Scale bar: 100 µm. D , G Representative images ( D ) and quantification ( G ) of cell migration assessed by a Transwell assay. Migrated cells were stained with crystal violet ( n = 5). Scale bar: 50 µm. All data are presented as the mean ± SEM. Statistical significance was determined by one-way ANOVA followed by Tukey's post-hoc test. * P < 0.05, **P < 0.01, ***P < 0.001, ns , not significant

    Article Snippet: After washing with PBS, the sections were incubated with a Cy3-conjugated anti-rabbit at RT for 1 h, followed by labeling the cell nuclei with DAPI (Servicebio).

    Techniques: Knockdown, Migration, Transfection, Negative Control, Cell Characterization, CCK-8 Assay, Wound Healing Assay, Transwell Assay, Staining

    circATXN2 acting as a miR-138-5p sponge. A RNA Immunoprecipitation (RIP) for AGO2. qRT-PCR shows significant enrichment of circATXN2 in AGO2 pulldowns compared to IgG controls in PASMCs ( n = 3). B Venn diagram of predicted miRNA targets for circATXN2 from miRanda, TargetScan, and RNAhybrid, identifying miR-138-5p as a top candidate. C RNA pulldown assay using biotinylated miR-138-5p or other candidate miRNAs identified in (B). qRT-PCR analysis shows specific and significant enrichment of circATXN2 only with the miR-138-5p probe ( n = 3). D Schematic of circATXN2 (318 nt) showing the 6 predicted binding sites for miR-138-5p. E Co-localization of circATXN2 (red) and miR-138-5p (green) in PASMCs via FISH assays, demonstrating co-localization in the cytoplasm. Nuclei are counterstained with DAPI (blue). Scale bar: 50 µm. F , G qRT-PCR analysis of miR-138-5p levels in PASMCs following (F) knockdown of circATXN2 (si-circATXN2) or (G) overexpression of circATXN2 (oe-circATXN2), demonstrating reciprocal regulation ( n = 3). H-M Functional rescue experiments. PASMCs were co-transfected with a miR-138-5p mimic or negative control (miR-NC), along with an overexpression vector for circATXN2 (oe-circATXN2) or its control (oe-NC). CCK-8 viability assay ( n = 5) (H). Representative images (I) and quantification (K) of EdU proliferation assay ( n = 3). Scale bar: 100 µm. Representative images (J) and quantification (M) of Transwell migration assay ( n = 3). Scale bar: 50 µm. Quantification of wound healing migration assay ( n = 3) (L). The miR-138-5p mimic reversed the pro-proliferative and pro-migratory effects of circATXN2 overexpression. All data are presented as the mean ± SEM. Statistical significance was determined using a two-tailed Student's t-test (for A, C, F, G) or one-way ANOVA followed by Tukey's post-hoc test (for H, K, L, M). * P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, ns , not significant

    Journal: Respiratory Research

    Article Title: CircATXN2 exacerbates pulmonary arterial hypertension by modulating the miR-138-5p/SMURF1 axis

    doi: 10.1186/s12931-026-03647-w

    Figure Lengend Snippet: circATXN2 acting as a miR-138-5p sponge. A RNA Immunoprecipitation (RIP) for AGO2. qRT-PCR shows significant enrichment of circATXN2 in AGO2 pulldowns compared to IgG controls in PASMCs ( n = 3). B Venn diagram of predicted miRNA targets for circATXN2 from miRanda, TargetScan, and RNAhybrid, identifying miR-138-5p as a top candidate. C RNA pulldown assay using biotinylated miR-138-5p or other candidate miRNAs identified in (B). qRT-PCR analysis shows specific and significant enrichment of circATXN2 only with the miR-138-5p probe ( n = 3). D Schematic of circATXN2 (318 nt) showing the 6 predicted binding sites for miR-138-5p. E Co-localization of circATXN2 (red) and miR-138-5p (green) in PASMCs via FISH assays, demonstrating co-localization in the cytoplasm. Nuclei are counterstained with DAPI (blue). Scale bar: 50 µm. F , G qRT-PCR analysis of miR-138-5p levels in PASMCs following (F) knockdown of circATXN2 (si-circATXN2) or (G) overexpression of circATXN2 (oe-circATXN2), demonstrating reciprocal regulation ( n = 3). H-M Functional rescue experiments. PASMCs were co-transfected with a miR-138-5p mimic or negative control (miR-NC), along with an overexpression vector for circATXN2 (oe-circATXN2) or its control (oe-NC). CCK-8 viability assay ( n = 5) (H). Representative images (I) and quantification (K) of EdU proliferation assay ( n = 3). Scale bar: 100 µm. Representative images (J) and quantification (M) of Transwell migration assay ( n = 3). Scale bar: 50 µm. Quantification of wound healing migration assay ( n = 3) (L). The miR-138-5p mimic reversed the pro-proliferative and pro-migratory effects of circATXN2 overexpression. All data are presented as the mean ± SEM. Statistical significance was determined using a two-tailed Student's t-test (for A, C, F, G) or one-way ANOVA followed by Tukey's post-hoc test (for H, K, L, M). * P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, ns , not significant

    Article Snippet: After washing with PBS, the sections were incubated with a Cy3-conjugated anti-rabbit at RT for 1 h, followed by labeling the cell nuclei with DAPI (Servicebio).

    Techniques: RNA Immunoprecipitation, Quantitative RT-PCR, Binding Assay, Knockdown, Over Expression, Functional Assay, Transfection, Negative Control, Plasmid Preparation, Control, CCK-8 Assay, Viability Assay, Proliferation Assay, Transwell Migration Assay, Migration, Two Tailed Test

    Reducing circATXN2 in SMCs attenuated the severity of PAH via miR-138-5p/SMURF1 axis. A Immunofluorescence staining of pulmonary arteries confirming SMC-specific AAV9 transduction. GFP (AAV9, green) co-localizes with the SMC marker α-SMA (red). Nuclei were counterstained with DAPI (blue). Scale bar: 100 µm. B qRT-PCR analysis of circATXN2 expression in lung tissue from mice injected with AAV9-si-NC or AAV9-si-circATXN2, confirming successful knockdown ( n = 6) . C Quantification of Right Ventricular Systolic Pressure (RVSP) in the four treatment groups ( n = 6). D , E Echocardiographic measurements of AT/ET ratio (D) and PAVTI (E) in the four treatment groups ( n = 6). F , G Quantification (F) and Hematoxylin and Eosin (H&E) staining (G) of pulmonary arterioles, showing reduced medial wall thickening in the AAV9-si-circATXN2-treated hypoxia group ( n = 6). Scale bar: 50 µm. H Western blot analysis of SMURF1 protein expression in lung lysates from the four treatment groups. β-actin was used as a loading control ( n = 6). I To validate the in vivo role of circATXN2, SMC-specific knockdown was achieved using AAV9-si-circATXN2 in 6-week-old Tagln-Cre mice. As shown in the schematic, mice were injected via the tail vein. After a two-week infection period, mice were exposed to normoxia (21% FiO 2 ) or chronic hypoxia (10% FiO 2 ) and weekly subcutaneous semaxanib (SU5416, 20 mg/kg/week) injections for 4 weeks to induce PAH. All data are presented as the mean ± SEM. Statistical significance was determined using a two-tailed Student's t-test (for B) or two-way ANOVA followed by Tukey's post-hoc test (for C-F). * P < 0.05, **P < 0.01, ns , not significant

    Journal: Respiratory Research

    Article Title: CircATXN2 exacerbates pulmonary arterial hypertension by modulating the miR-138-5p/SMURF1 axis

    doi: 10.1186/s12931-026-03647-w

    Figure Lengend Snippet: Reducing circATXN2 in SMCs attenuated the severity of PAH via miR-138-5p/SMURF1 axis. A Immunofluorescence staining of pulmonary arteries confirming SMC-specific AAV9 transduction. GFP (AAV9, green) co-localizes with the SMC marker α-SMA (red). Nuclei were counterstained with DAPI (blue). Scale bar: 100 µm. B qRT-PCR analysis of circATXN2 expression in lung tissue from mice injected with AAV9-si-NC or AAV9-si-circATXN2, confirming successful knockdown ( n = 6) . C Quantification of Right Ventricular Systolic Pressure (RVSP) in the four treatment groups ( n = 6). D , E Echocardiographic measurements of AT/ET ratio (D) and PAVTI (E) in the four treatment groups ( n = 6). F , G Quantification (F) and Hematoxylin and Eosin (H&E) staining (G) of pulmonary arterioles, showing reduced medial wall thickening in the AAV9-si-circATXN2-treated hypoxia group ( n = 6). Scale bar: 50 µm. H Western blot analysis of SMURF1 protein expression in lung lysates from the four treatment groups. β-actin was used as a loading control ( n = 6). I To validate the in vivo role of circATXN2, SMC-specific knockdown was achieved using AAV9-si-circATXN2 in 6-week-old Tagln-Cre mice. As shown in the schematic, mice were injected via the tail vein. After a two-week infection period, mice were exposed to normoxia (21% FiO 2 ) or chronic hypoxia (10% FiO 2 ) and weekly subcutaneous semaxanib (SU5416, 20 mg/kg/week) injections for 4 weeks to induce PAH. All data are presented as the mean ± SEM. Statistical significance was determined using a two-tailed Student's t-test (for B) or two-way ANOVA followed by Tukey's post-hoc test (for C-F). * P < 0.05, **P < 0.01, ns , not significant

    Article Snippet: After washing with PBS, the sections were incubated with a Cy3-conjugated anti-rabbit at RT for 1 h, followed by labeling the cell nuclei with DAPI (Servicebio).

    Techniques: Immunofluorescence, Staining, Transduction, Marker, Quantitative RT-PCR, Expressing, Injection, Knockdown, Western Blot, Control, In Vivo, Infection, Two Tailed Test