rabbit anti cdc25a  (Cell Signaling Technology Inc)

Journal: Respiratory Research

Article Title: The mechanism by which KAT2A increases the stability of CDC25A through acetylation to regulate glycolysis and mediate lung adenocarcinoma immune escape

doi: 10.1186/s12931-026-03544-2

Figure Lengend Snippet: Upregulation of CDC25A in LUAD tissues and cells. A The TCGA-LUAD database provides an analysis of CDC25A expression, where blue corresponds to normal tissues and red to tumor tissues; ( B ) Kaplan-Meier analysis of CDC25A in LUAD patients; ( C ) qRT-PCR detection of CDC25A mRNA in human lung epithelial cells (BEAS-2B) and LUAD cell lines (A549, H1299, Calu-3); ( D ) qRT-PCR detection of CDC25A mRNA in 12 pairs of LUAD and adjacent normal tissues; ( E ) WB analysis of CDC25A protein expression levels in human lung epithelial cells (BEAS-2B) and LUAD cell lines (A549, H1299, Calu-3). * represents P < 0.05

Article Snippet: Cells were permeabilized with 0.5% Triton X-100 (Sigma, USA) for 10 min at room temperature, then blocked with 5% BSA for 1 h. Primary antibodies specific for CDC25A (A1173, ABClonal, China) and HK2 (ab209847, Abcam, USA) were applied and incubated overnight.

Techniques: Expressing, Quantitative RT-PCR

Journal: Respiratory Research

Article Title: The mechanism by which KAT2A increases the stability of CDC25A through acetylation to regulate glycolysis and mediate lung adenocarcinoma immune escape

doi: 10.1186/s12931-026-03544-2

Figure Lengend Snippet: CDC25A knockdown enhances the cytotoxicity of CD8 + T cells against LUAD cells. A - B qRT-PCR and WB analysis of transfection efficiency for sh-NC and sh-CDC25A plasmids; ( C ) CCK-8 test for cell viability; ( D ) Live/death cell staining assay for detecting cell live and death; ( E ) Colony formation assay for evaluating cell proliferation; ( F ) Analysis of the correlation between CDC25A and CD8 + T cell immune infiltration by combining TCGA database and Timer database; ( G ) Flow cytometry detection of IFN-γ fluorescence intensity; ( H ) LDH detection of the cytotoxic effect of CD8 + T cells on LUAD cells; ( I ) ELISA for quantifying TNF-α, IFN-γ, Perforin, and Granzyme B in the co-culture supernatants. * represents P < 0.05

Article Snippet: Cells were permeabilized with 0.5% Triton X-100 (Sigma, USA) for 10 min at room temperature, then blocked with 5% BSA for 1 h. Primary antibodies specific for CDC25A (A1173, ABClonal, China) and HK2 (ab209847, Abcam, USA) were applied and incubated overnight.

Techniques: Knockdown, Quantitative RT-PCR, Transfection, CCK-8 Assay, Staining, Colony Assay, Flow Cytometry, Fluorescence, Enzyme-linked Immunosorbent Assay, Co-Culture Assay

Journal: Respiratory Research

Article Title: The mechanism by which KAT2A increases the stability of CDC25A through acetylation to regulate glycolysis and mediate lung adenocarcinoma immune escape

doi: 10.1186/s12931-026-03544-2

Figure Lengend Snippet: CDC25A enhances glycolysis to suppress the cytotoxicity of CD8 + T cells against LUAD cells. A - B ECAR and OCR levels of the two batches of culture from the same cell line were measured using the Seahorse XFe96 cell metabolic analyzer; ( C - D ) Glucose and lactate content were assessed with respective assay kits; ( E ) Cell viability was determined by CCK-8; ( F ) Live/death cell staining assay for detecting cell live and death; ( G ) Colony formation assay for cell proliferation capacity; ( H - I ) Flow cytometry for the activation level of CD8 + T cells; ( J ) LDH assay for the cytotoxic effect of CD8 + T cells on A549 cells; ( K ) ELISA for the content of TNF-α, IFN-γ, Perforin, and Granzyme B in co-culture supernatants. * represents P < 0.05

Article Snippet: Cells were permeabilized with 0.5% Triton X-100 (Sigma, USA) for 10 min at room temperature, then blocked with 5% BSA for 1 h. Primary antibodies specific for CDC25A (A1173, ABClonal, China) and HK2 (ab209847, Abcam, USA) were applied and incubated overnight.

Techniques: CCK-8 Assay, Staining, Colony Assay, Flow Cytometry, Activation Assay, Lactate Dehydrogenase Assay, Enzyme-linked Immunosorbent Assay, Co-Culture Assay

Journal: Respiratory Research

Article Title: The mechanism by which KAT2A increases the stability of CDC25A through acetylation to regulate glycolysis and mediate lung adenocarcinoma immune escape

doi: 10.1186/s12931-026-03544-2

Figure Lengend Snippet: CDC25A binds to HK2 to promote aerobic glycolysis in LUAD cells. A Molecular docking simulation of CDC25A and HK2 conducted with InterEvDock2, red: HK2, blue: CDC25A; ( B ) CO-IP detection of the interaction between CDC25A and HK2; ( C ) Cellular IF to assess the localization of CDC25A and HK2 within cells; ( D - E ) WB analysis of HK2 protein expression in three groups of cells; ( F - H ) Seahorse XFe96 cell metabolic analyzer used to measure ECAR and OCR levels in the three cell groups; ( I - J ) Glucose content and lactate production levels detected with respective assay kits. * represents P < 0.05

Article Snippet: Cells were permeabilized with 0.5% Triton X-100 (Sigma, USA) for 10 min at room temperature, then blocked with 5% BSA for 1 h. Primary antibodies specific for CDC25A (A1173, ABClonal, China) and HK2 (ab209847, Abcam, USA) were applied and incubated overnight.

Techniques: Co-Immunoprecipitation Assay, Expressing

Journal: Respiratory Research

Article Title: The mechanism by which KAT2A increases the stability of CDC25A through acetylation to regulate glycolysis and mediate lung adenocarcinoma immune escape

doi: 10.1186/s12931-026-03544-2

Figure Lengend Snippet: KAT2A mediates acetylation modification of H3K27ac to enhance CDC25A expression. A Analysis of KAT2A expression in LUAD tissues through the TCGA-LUAD database, with blue signifying normal tissue samples and red signifying tumor tissue samples; ( B ) Pearson correlation analysis between CDC25A and KAT2A; ( C ) qRT-PCR for KAT2A mRNA levels in human pulmonary epithelial cells and LUAD cell lines; ( D - E ) WB analysis of KAT2A protein expression levels in human pulmonary epithelial cells and LUAD cell lines; ( F ) ChIP experiment confirming the interaction between CDC25A and KAT2A; ( G ) qRT-PCR evaluation of transfection efficiency of sh-NC/sh-KAT2A plasmids; ( H ) qRT-PCR analysis of CDC25A mRNA expression levels after KAT2A knockdown; ( I ) WB analysis of KAT2A and CDC25A protein expression levels after KAT2A knockdown; ( J ) ChIP experiment verifying H3K27ac modification of CDC25A mRNA in A549 cells; ( K ) qRT-PCR detection of CDC25A mRNA expression following KAT2A knockdown. * represents P < 0.05

Article Snippet: Cells were permeabilized with 0.5% Triton X-100 (Sigma, USA) for 10 min at room temperature, then blocked with 5% BSA for 1 h. Primary antibodies specific for CDC25A (A1173, ABClonal, China) and HK2 (ab209847, Abcam, USA) were applied and incubated overnight.

Techniques: Modification, Expressing, Quantitative RT-PCR, Transfection, Knockdown

Journal: Respiratory Research

Article Title: The mechanism by which KAT2A increases the stability of CDC25A through acetylation to regulate glycolysis and mediate lung adenocarcinoma immune escape

doi: 10.1186/s12931-026-03544-2

Figure Lengend Snippet: KAT2A inhibits the cytotoxic capacity of CD8 + T cells against LUAD cells by interacting with CDC25A. A - B qRT-PCR and WB analysis of CDC25A mRNA expression levels in the three cell groups; ( C ) CCK-8 assay for cell viability assessment; ( D ) Live/death cell staining assay for detecting cell live and death; ( E ) Colony formation assay for cell proliferation capacity; ( F - G ) Seahorse XFe96 cell metabolic analyzer for ECAR and OCR levels in the three cell groups; ( H - I ) Glucose content and lactate production measured by respective assay kits; ( J ) Flow cytometry for CD8 + T cell activation levels in the three groups; ( K ) LDH assay for the cytotoxic effect of CD8 + T cells on A549 cells; ( L ) ELISA for TNF-α, IFN-γ, Perforin, and Granzyme B levels in co-culture supernatants. * represents P < 0.05

Article Snippet: Cells were permeabilized with 0.5% Triton X-100 (Sigma, USA) for 10 min at room temperature, then blocked with 5% BSA for 1 h. Primary antibodies specific for CDC25A (A1173, ABClonal, China) and HK2 (ab209847, Abcam, USA) were applied and incubated overnight.

Techniques: Quantitative RT-PCR, Expressing, CCK-8 Assay, Staining, Colony Assay, Flow Cytometry, Activation Assay, Lactate Dehydrogenase Assay, Enzyme-linked Immunosorbent Assay, Co-Culture Assay

Journal: Respiratory Research

Article Title: The mechanism by which KAT2A increases the stability of CDC25A through acetylation to regulate glycolysis and mediate lung adenocarcinoma immune escape

doi: 10.1186/s12931-026-03544-2

Figure Lengend Snippet: In vivo validation of the regulatory role of the KAT2A/CDC25A/HK2 axis on tumor growth and immune status. A - C Documentation of tumor growth in murine models; ( D ) IHC analysis of CDC25A, KAT2A, and HK2 expression levels in tumor tissues; ( E ) WB analysis of CDC25A and KAT2A expression levels in tumor tissues; ( F ) IF analysis of CD8 + T cell infiltration in tumor tissues; ( G ) ELISA for TNF-α, IFN-γ, Perforin, and Granzyme B levels in peripheral blood. * represents P < 0.05

Article Snippet: Cells were permeabilized with 0.5% Triton X-100 (Sigma, USA) for 10 min at room temperature, then blocked with 5% BSA for 1 h. Primary antibodies specific for CDC25A (A1173, ABClonal, China) and HK2 (ab209847, Abcam, USA) were applied and incubated overnight.

Techniques: In Vivo, Biomarker Discovery, Expressing, Enzyme-linked Immunosorbent Assay

Journal: PLOS ONE

Article Title: Cyclosporine A causes gingival overgrowth via reduced G 1 cell cycle arrest in gingival fibroblasts

doi: 10.1371/journal.pone.0309189

Figure Lengend Snippet: Primers used for reverse transcription-quantitative PCR.

Article Snippet: The rabbit anti-CDC25A antibody (cat. no. 55031-1-AP) was purchased from Proteintech Group, Inc (Rosemont, IL, USA).

Techniques: Sequencing

Journal: PLOS ONE

Article Title: Cyclosporine A causes gingival overgrowth via reduced G 1 cell cycle arrest in gingival fibroblasts

doi: 10.1371/journal.pone.0309189

Figure Lengend Snippet: Semiconfluent cells were incubated in D-MEM containing 2% serum with or without (control) 200 ng/mL (166 nM) CsA for 12 h, after which reverse transcription-quantitative PCR analysis was performed. Relative quantification was performed using the 2‑ΔΔCq method. After normalization to GAPDH, RNA ratios in treated vs. control cultures were determined. Data are presented as means ± SEM. * P <0.05 compared with the control using Welch’s t-test (n = 3). (A) G 1 -S transition-promoted genes ( CDC25A , cell division cycle 25A; CDK2 , cyclin dependent kinase 2; CYCLIN E1 , E-type cyclin; MYC , MYC proto-oncogene, bHLH transcription factor). (B) G 1 arrest-induced genes ( ATM , ATM serine/threonine kinase; ATR , ATR serine/threonine kinase; P21 , cyclin dependent kinase inhibitor 1A; P27 , cyclin dependent kinase inhibitor 1B; P53 , tumor protein p53; RB1 , RB transcriptional corepressor 1; SMAD3 , SMAD family member 3; SMAD4 , SMAD family member 4).

Article Snippet: The rabbit anti-CDC25A antibody (cat. no. 55031-1-AP) was purchased from Proteintech Group, Inc (Rosemont, IL, USA).

Techniques: Incubation, Control, Reverse Transcription, Real-time Polymerase Chain Reaction, Quantitative Proteomics

Journal: PLOS ONE

Article Title: Cyclosporine A causes gingival overgrowth via reduced G 1 cell cycle arrest in gingival fibroblasts

doi: 10.1371/journal.pone.0309189

Figure Lengend Snippet: Semiconfluent cells were incubated in D-MEM containing 2% serum with or without (control) 200 ng/mL (166 nM) CsA for 24 h and were then assessed using western blotting, after which the fold change compared with the control was determined. The band images shown are representative of results from three independent experiments. Data are presented as means ± SEM. * P <0.05 compared with the control using Welch’s t-test (n = 3). (A) G 1 -S transition-promoted proteins (CDC25A, cell division cycle 25A; CYCLIN E1, E-type cyclin; pCDK2, phospho-cyclin dependent kinase 2; RB1, RB transcriptional corepressor 1). (B) G 1 arrest-induced proteins (P21, cyclin dependent kinase inhibitor 1A; SMAD3, SMAD family member 3; SMAD4, SMAD family member 4).

Article Snippet: The rabbit anti-CDC25A antibody (cat. no. 55031-1-AP) was purchased from Proteintech Group, Inc (Rosemont, IL, USA).

Techniques: Incubation, Control, Western Blot

Journal: PLOS ONE

Article Title: Cyclosporine A causes gingival overgrowth via reduced G 1 cell cycle arrest in gingival fibroblasts

doi: 10.1371/journal.pone.0309189

Figure Lengend Snippet: CsA induces the downregulation of SMAD3 and SMAD4, which leads to a decrease in P21. CsA also induces the upregulation of CDC25A, CYCLIN E, phosphorylated CDK2 and phosphorylated RB, which results in the inhibition of G 1 cell cycle arrest. Light blue (factors of G 1 arrest-inducing signaling) and light green (factors of G 1 /S transition-inducing signaling) rectangles similarly denote the molecules analyzed in this study. The blue or red large arrows denote downregulation or upregulation, respectively, following CsA treatment. ATM, ATM serine/threonine kinase; ATR, ATR serine/threonine kinase; CDC25A, cell division cycle 25A; CYCLIN E, E-type cyclin; MYC, MYC proto-oncogene, bHLH transcription factor; pCDK2, phosphorylated cyclin dependent kinase 2; pRB1, phosphorylated RB transcriptional corepressor 1; P21, cyclin dependent kinase inhibitor 1A; P27, cyclin dependent kinase inhibitor 1B; P53, tumor protein p53; RB1, RB transcriptional corepressor 1; SMAD3, SMAD family member 3; SMAD4, SMAD family member 4.

Article Snippet: The rabbit anti-CDC25A antibody (cat. no. 55031-1-AP) was purchased from Proteintech Group, Inc (Rosemont, IL, USA).

Techniques: Inhibition