Journal: Journal for Immunotherapy of Cancer

Article Title: Anti-CTGF/PD-1 bispecific antibody Y126S restrains desmoplastic and immunosuppressive microenvironment in pancreatic cancer

doi: 10.1136/jitc-2025-012144

Figure Lengend Snippet: The basic characteristics of CAFs-targeting anti-CTGF/PD-1 bispecific antibody (Y126S). ( A ) Y126S was designed with a structure comprizing two anti-CTGF regions and two anti-PD-1 regions. Schematic representation of Y126S bispecific binding via CTGF and PD-1 in the tumor site. ( B ) The results of non-reduced and reduced SDS-PAGE assay. A single band was found in the lane of non-reduced Y126S monomers, and two bands were observed in the lane of Y126S. ( C–H ) Binding affinity to hCTGF, mCTGF, and mPD-1 for neutralizing antibodies at the indicated concentrations was assessed by surface plasmon resonance. ( C, D ) Binding affinity to hCTGF antigen for Y126S and α-CTGF at the indicated concentrations by surface plasmon resonance. ( E, F ) Binding affinity to mCTGF antigen for Y126S and α-CTGF at the indicated concentrations by surface plasmon resonance. ( G, H ) Binding affinity to mPD-1 antigen for Y126S and α-PD-1 at the indicated concentrations by surface plasmon resonance. ( I, J ) Indirect anti-hIgG ELISA showed the binding affinity of serially diluted Y126S, binding controls, or negative controls to mCTGF and mPD-1. ( K ) Double-antigen sandwich ELISA showed that Y126S simultaneously bound mCTGF and mPD-1 at indicated concentrations. n = 3 replicates in I–K. CAF, cancer-associated fibroblast; CTGF, connective tissue growth factor; hCTGF, human CTGF antigen; mCTGF, mouse CTGF antigen; mPD-1, mouse programmed cell death 1 antigen; OD, optimal density; PD-L1, programmed cell death ligand 1; SDS-PAGE, Sodium dodecyl sulfate-polyacrylamide gel electrophoresis.

Article Snippet: Three replicates of hCAFs isolated from patients with PDAC were treated with recombinant human CTGF (rhCTGF) (HY- P70106 , MedChemExpress) for 2 days.

Techniques: Binding Assay, SDS Page, SPR Assay, Enzyme-linked Immunosorbent Assay, Sandwich ELISA, Polyacrylamide Gel Electrophoresis

Journal: Journal for Immunotherapy of Cancer

Article Title: Anti-CTGF/PD-1 bispecific antibody Y126S restrains desmoplastic and immunosuppressive microenvironment in pancreatic cancer

doi: 10.1136/jitc-2025-012144

Figure Lengend Snippet: Y126S reversed CTGF-mediated immunosuppressive TME. ( A ) Correlation analysis of CTGF with PD-L1 expression and exhausted CD8 + T cells in TCGA database (n=179). ( B ) Gene set enrichment analysis (GSEA) correlates CAFs-CTGF function with T cell differentiation, immune response regulation, and immune response regulating cell surface receptor signaling pathway in TCGA database (n=179). ( C, D ) Primary hCAFs isolated from patients with PDAC were treated with 20 ng/mL rhCTGF for 48 hours and subjected to RNA-seq analysis. Heatmap of differentially expressed genes ( C ) and top pathways in GO enrichment ( D ). ( E, F ) PD-L1 expression of hCAFs ( E ) and KPC-CAFs ( F ) after treatment with CM from cancer cells or CTGF (20 ng/mL) with α-CTGF or Y126S for 48 hours. ( G–P ) The effects of α-CTGF injection in orthotopic PDAC-bearing mice were evaluated. Schematic illustration for the design ( G ), bioluminescence images and average radiance ( H ), tumor weight ( I ), and representative images and correlation analysis between CTGF and PD-L1 expression in IHC staining ( J ). Quantification for the percentage of CD3 + T cells ( K ), CD8 + T cells ( L ), PD-1 + TIM-3 + T cells in CD8 + T cells ( M ), and expression of CD206 + by TAMs ( N ) of tumors treated with α-CTGF by flow cytometry. Representative images and quantification of anti-CD8 ( O ) and anti-Ki67 ( P ) IHC staining in tumors treated with α-CTGF or IgG group. 7 mice per group (n = 7) in G–P. Data were represented as mean ± SEM. Statistical analysis was performed with two-tailed Student’s t-tests. * p < 0.05, ** p < 0.01, *** p < 0.001, and ns = not significant. Scale bar: 20 µm. AOD, average optimal density; CM, conditioned media; CTGF, connective tissue growth factor; FACS, fluorescence-activated cell sorting; FDR, false discovery rate; GO, gene oncology; hCAFs, human cancer-associated fibroblasts; IHC, immunohistochemistry; IVIS, In Vivo Imaging System; KPC-CAFs, KPC’s cancer-associated fibroblasts; mCTGF, mouse CTGF protein; NES, nonparametric estimation statistics; PDAC, pancreatic ductal adenocarcinoma; PD-L1, programmed cell death ligand 1; rhCTGF, recombinant human CTGF protein; TAMs, tumor-associated macrophages; TCGA, The Cancer Genome Atlas; TME, tumor microenvironment.

Article Snippet: Three replicates of hCAFs isolated from patients with PDAC were treated with recombinant human CTGF (rhCTGF) (HY- P70106 , MedChemExpress) for 2 days.

Techniques: Expressing, Cell Differentiation, Cell Surface Receptor Assay, Isolation, RNA Sequencing, Injection, Immunohistochemistry, Flow Cytometry, Two Tailed Test, Fluorescence, FACS, In Vivo Imaging, Recombinant

Journal: Journal for Immunotherapy of Cancer

Article Title: Anti-CTGF/PD-1 bispecific antibody Y126S restrains desmoplastic and immunosuppressive microenvironment in pancreatic cancer

doi: 10.1136/jitc-2025-012144

Figure Lengend Snippet: Y126S counteracts CTGF-caused immunosuppression by downregulating PD-L1 expression and reactivating T cell immune response. ( A ) KPC-CAFs were pretreated with the stimulation of 20 ng/mL mCTGF plus multiple antibodies for 48 hours and then co-cultured with CD8 + T cells isolated from C57BL/6J spleen for 18 hours (effector/target ratio = 2:1). Representative flow cytometric images and corresponding quantifications of PD-L1 expression by KPC-CAFs (top), ratio of PD-1 + TIM-3 + T cells in CD8 + T cells (center), and apoptosis of CD8 + T cells by flow cytometry (bottom). ( B ) Level of IL-2, IFN-γ, and TNF-α secreted by T cells in a co-culture system. 3 replicates per group (n = 3) in all experiments. Data were represented as mean ± SEM. Statistical analysis was performed with ANOVA followed by Tukey’s multiple comparison test. * p < 0.05, ** p < 0.01, *** p < 0.001, and ns = not significant. ANOVA, analysis of variance; comb, combination of α-CTGF and α-PD-1; IFN-γ, interferon-gamma; KPC-CAFs, KPC’s cancer-associated fibroblasts; mCTGF, mouse connective tissue growth factor protein; PD-1, programmed cell death 1; PD-L1, programmed cell death ligand 1; TNF-α, tumor necrosis factor-alpha.

Article Snippet: Three replicates of hCAFs isolated from patients with PDAC were treated with recombinant human CTGF (rhCTGF) (HY- P70106 , MedChemExpress) for 2 days.

Techniques: Expressing, Cell Culture, Isolation, Flow Cytometry, Co-Culture Assay, Comparison

Journal: Journal for Immunotherapy of Cancer

Article Title: Anti-CTGF/PD-1 bispecific antibody Y126S restrains desmoplastic and immunosuppressive microenvironment in pancreatic cancer

doi: 10.1136/jitc-2025-012144

Figure Lengend Snippet: Y126S inhibited desmoplasia induced by CAF activation through targeting CTGF. ( A–E ) The desmoplasia of tumors derived from PDAC mice treated with α-CTGF or IgG in . Representative images ( A ), quantification of anti-α-SMA ( B ), anti-collagen I ( C ), Masson ( D ) staining, and tissue stiffness ( E ). ( F, G ) Relative expression of specific genes of hCAFs after treatment with CM from PANC-1 cells or rhCTGF (20 ng/mL) plus α-CTGF or Y126S for 48 hours by qRT-PCR ( F ) and western blotting ( G ). ( H ) Representative images of hCAFs stimulated by rhCTGF (20 ng/mL) plus α-CTGF or Y126S for 48 hours for α-SMA (green), collagen I (red), and DAPI (blue). ( I, J ) Relative expression of specific genes of KPC-CAFs after treatment with CM from Panc-02 cells or mCTGF (20 ng/mL) plus α-CTGF or Y126S for 48 hours by qRT-PCR ( I ) and western blotting ( J ). ( K–M ) Representative fast force MAP AFM image ( K ), Young’s modulus of hCAFs ( L ), and KPC-CAFs ( M ) by AFM with a CSC38 cantilever (spring constant: 0.02–0.04 N/m). ( N ) Gel contraction of hCAFs treated with 20 ng/mL rhCTGF plus α-CTGF or Y126S. 7 replicates per group (n = 7) in B–E, 3 replicates per group (n = 3) in F, J, N, and 5 replicates per group (n = 5) in L, M. Data were represented as mean ± SEM. Statistical analysis in B–E was performed with two-tailed Student’s t-tests, and in F, J, L–N was performed with ANOVA followed by Tukey’s multiple comparison test. * p < 0.05, ** p < 0.01, *** p < 0.001, ns = not significant. Scale bar: 20 µm in A, 50 µm in H, and 10 µm in K. AFM, atomic force microscopy; ANOVA, analysis of variance; α-SMA, alpha-smooth muscle actin; CM, conditioned media; CTGF, connective tissue growth factor; DAPI, 4',6-diamidino-2-phenylindole; FAP, fibroblast activation protein; hCAFs, human cancer-associated fibroblasts; KPC-CAFs, KPC’s cancer-associated fibroblasts; mCTGF, mouse CTGF protein; mRNA, messenger RNA; PDAC, pancreatic ductal adenocarcinoma; qRT-PCR, quantitative reverse transcription PCR; rhCTGF, recombinant human CTGF protein.

Article Snippet: Three replicates of hCAFs isolated from patients with PDAC were treated with recombinant human CTGF (rhCTGF) (HY- P70106 , MedChemExpress) for 2 days.

Techniques: Activation Assay, Derivative Assay, Staining, Expressing, Quantitative RT-PCR, Western Blot, Two Tailed Test, Comparison, Microscopy, Reverse Transcription, Recombinant

Journal: Journal for Immunotherapy of Cancer

Article Title: Anti-CTGF/PD-1 bispecific antibody Y126S restrains desmoplastic and immunosuppressive microenvironment in pancreatic cancer

doi: 10.1136/jitc-2025-012144

Figure Lengend Snippet: Y126S exhibits potent antitumor efficiency in PDAC mouse model by remodeling TME. ( A–C ) The effects of Y126S with different dosages in orthotopic PDAC mice. Schematic illustration for the design ( A ), tumor weight ( B ), and survival curve ( C ) of PDAC-bearing mice after receiving IgG, low (4 mg/kg), middle (8 mg/kg), and high dose (16 mg/kg) of Y126S treatment. ( D–Q ) The effects of IgG, α-CTGF, α-PD-1, a combination of monospecific antibodies, and Y126S in orthotopic PDAC mice were evaluated. Schematic illustration ( D ), bioluminescence images ( E ), average radiance ( F ), and tumor weight ( G ) of mice after the antibody treatment as indicated. Representative images ( H ), quantification of anti-α-SMA ( I ), anti-collagen I ( J ), Masson ( K ) staining, and tissue stiffness ( L ) in PDAC tumors. Proportion of CD3 + T cells ( M ) and CD8 + T cells ( N ) in live cells, PD-1 + TIM-3 + T cells in CD8 + T cells ( O ), and expression of CD206 + by CD11b + F4/80 + TAMs ( P ) in tumors of different treatment groups by flow cytometry. ( Q ) The level of IFN-γ in tumor tissues by ELISA assay. 8 mice per group (n = 8) in all experiments. Data were represented as mean ± SEM. Statistical analysis was performed with ANOVA followed by Tukey’s multiple comparison test. * p < 0.05, ** p < 0.01, *** p < 0.001, and ns = not significant. Scale bar: 20 µm. ANOVA, analysis of variance; α-SMA, alpha smooth muscle actin; comb, combination of α-CTGF and α-PD-1; CTGF, connective tissue growth factor; IFN-γ, interferon-gamma; i.p., intraperitoneal; IVIS, In Vivo Imaging System; PD-1, programmed cell death 1; PDAC, pancreatic ductal adenocarcinoma; TAMs, tumor-associated macrophages; TME, tumor microenvironment.

Article Snippet: Three replicates of hCAFs isolated from patients with PDAC were treated with recombinant human CTGF (rhCTGF) (HY- P70106 , MedChemExpress) for 2 days.

Techniques: Staining, Expressing, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Comparison, In Vivo Imaging

Journal: Journal for Immunotherapy of Cancer

Article Title: Anti-CTGF/PD-1 bispecific antibody Y126S restrains desmoplastic and immunosuppressive microenvironment in pancreatic cancer

doi: 10.1136/jitc-2025-012144

Figure Lengend Snippet: Y126S yields a robust antitumor response and prolongs survival in the KPC model. The effects of IgG, α-CTGF, α-PD-1, a combination of monospecific antibodies, and Y126S were validated in the KPC mouse model. ( A ) Schematic illustration of study design. ( B ) 68 Ga-FAPI PET MIP images of different groups before and after treatment. The white dotted circles and red arrows indicate the tumor sites. ΔSUVmax of each tumor was analyzed by subtracting the SUVmax measured pretreatment from the SUVmax after treatment. ( C, D ) The tumor weight ( C ) and overall survival ( D ) of KPC mice in different groups. ( E–N ) Tumors were then further evaluated to understand the change in TME of KPC mice. Representative images ( E ) and quantification of anti-α-SMA ( F ), anti-collagen I ( G ), Masson ( H ) staining, and tissue stiffness ( I ). The proportion of CD3 + T cells ( J ) and CD8 + T cells ( K ) in live cells, PD-1 + TIM-3 + T cells in CD8 + T cells ( L ), and expression of CD206 + by CD11b + F4/80 + TAMs ( M ) by flow cytometry. ( N ) The level of IFN-γ in tumor tissues by ELISA assay. 3 mice in IgG, α-CTGF, α-PD-1 group, 4 in the combination group, and 6 in Y126S group in B due to survival statement in D, 6 mice per group (n = 6) in C, F–N. Data were represented as mean ± SEM. Statistical analysis was performed with ANOVA followed by Tukey’s multiple comparison test. * p < 0.05, ** p < 0.01, *** p < 0.001. Scale bar: 20 µm. ANOVA, analysis of variance; α-SMA, alpha-smooth muscle actin; comb, a combination of α-CTGF and α-PD-1; CTGF, connective tissue growth factor; IFN-γ, interferon-gamma; i.p., intraperitoneal; PD-1, programmed cell death 1; PET, positron emission tomography; SUVmax, maximum of standard uptake values; TAMs, tumor-associated macrophages; TME, tumor microenvironment.

Article Snippet: Three replicates of hCAFs isolated from patients with PDAC were treated with recombinant human CTGF (rhCTGF) (HY- P70106 , MedChemExpress) for 2 days.

Techniques: Staining, Expressing, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Comparison, Positron Emission Tomography

Journal: Journal for Immunotherapy of Cancer

Article Title: Anti-CTGF/PD-1 bispecific antibody Y126S restrains desmoplastic and immunosuppressive microenvironment in pancreatic cancer

doi: 10.1136/jitc-2025-012144

Figure Lengend Snippet: Y126S exhibits tumor-specific accumulation through targeting CTGF. ( A–C ) The expression of CTGF in tumor tissues and adjacent normal tissue from patients with PDAC by RT-qPCR ( A ), western blotting ( B ), and IHC staining ( C ). For α-CTGF staining in C, representative histological plots were captured both in 100× and 200× (left panel), and quantification was conducted by measuring the positive area percentage of CTGF. n=5 (individual donors, illustrated as three technical replicates) (right panel). ( D ) Schematic illustration of the procedure of mice sacrifices and sample collection over time (0.5 hour, 1 hour, 2 hours, 4 hours, 8 hours, 24 hours, and 48 hours) after receiving a single administration of Y126S or α-PD-1. ( E ) Concentration curve of Y126S in serum. ( F, G ) Molecular weight-normalized-tumor concentration of Y126S and α-PD-1 in orthotopic tumor-bearing mice over time ( F ) and at 2 hours ( G ), expressed as molar concentration. ( H–K ) Further mass-normalized antibody levels of Y126S in tumor compared with liver ( H ) or lung ( I ), and of α-PD-1 in tumor compared with liver ( J ) or lung ( K ) were expressed as the percentage of molar concentration per gram of tissue. ( L ) Schematic illustration depicting the mechanism of Y126S treatment repressing desmoplastic and immunosuppressive TME in PDAC (created in BioRender.com). 3 replicates per group (n = 3) in A, 3 mice per group at each time point (n = 3) in D–K. Data were represented as mean ± SEM. Statistical analysis was performed with two-tailed Student’s t-tests. * p < 0.05, ** p < 0.01, *** p < 0.001. CAF, cancer-associated fibroblast; CTGF, connective tissue growth factor; ECM, extracellular matrix; IHC, immunohistochemistry; mRNA, messenger RNA; PD-1, programmed cell death 1; PDAC, pancreatic ductal adenocarcinoma; PD-L1, programmed cell death ligand 1; RT-qPCR, reverse transcription quantitative PCR; TAM, tumor-associated macrophage; TME, tumor microenvironment.

Article Snippet: Three replicates of hCAFs isolated from patients with PDAC were treated with recombinant human CTGF (rhCTGF) (HY- P70106 , MedChemExpress) for 2 days.

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Immunohistochemistry, Staining, Concentration Assay, Molecular Weight, Two Tailed Test, Reverse Transcription, Real-time Polymerase Chain Reaction

Journal: Journal of pharmacological sciences

Article Title: Oligo-peptide I-C-F-6 inhibits hepatic stellate cell activation and ameliorates CCl 4 -induced liver fibrosis by suppressing NF-κB signaling and Wnt/β-catenin signaling.

doi: 10.1016/j.jphs.2018.01.003

Figure Lengend Snippet: Fig. 3. The effects of oligo-peptide I-C-F-6 on HSC activation markers and cytokine levels. (A) The immunohistochemistry of TGF-b1 and a-SMA. Magnification, 200. (B) The effects of oligo-peptide I-C-F-6 on TGF-b1 and a-SMA levels in vitro. The data are expressed as the means ± SD of three independent experiments. *P < 0.05, **P < 0.01 compared with 0 mg/ mL. (C) The effects of oligo-peptide I-C-F-6 on TGF-b1 and a-SMA levels in vivo. (DeG) Serum levels of CTGF, TNF-a, VEGF, TGF-b1. Data are expressed as means ± SD (n ¼ 6e8). #P < 0.05, ##P < 0.01 compared with the control group; *P < 0.05, **P < 0.01 compared with the CCl4 group.

Article Snippet: The MMP-2, MMP-9, TIPM-1, CTGF, TGF-b1, TNF-a, and VEGF ELISA kits were purchased from Cusabio Biotech (Wuhan, China).

Techniques: Activation Assay, Immunohistochemistry, In Vitro, In Vivo, Control

Journal: STAR Protocols

Article Title: Generation and differentiation of chemically derived hepatic progenitors from mouse primary hepatocytes

doi: 10.1016/j.xpro.2021.100840

Figure Lengend Snippet:

Article Snippet: PAS stain kit , Abcam , Cat#ab150680.

Techniques: Recombinant, Staining, Software

Journal: Journal of Tissue Engineering

Article Title: Therapeutic potential and mechanisms of umbilical cord mesenchymal stem cells differentiating into tendon cells and promotion of rotator cuff tendon-bone healing

doi: 10.1177/20417314251315185

Figure Lengend Snippet: Analysis of the induction effect of BMP-12, GDF-7, and CTGF on the differentiation of UCMSCs: (a) morphological changes of UCMSCs after induction treatment, (b) mRNA expression of MKX, SCX, and TNC detected by qPCR, (c) bands and protein expression of MKX, SCX, and TNC detected by WB, and (d) COL-I and COL-III expression detected by immunofluorescence (400×). ** p < 0.01 and *** p < 0.001 compared with the control group.

Article Snippet: Rat umbilical cord mesenchymal stem cells (UCMSCs, CP-R302, Pricella, China), 293T cells (Pricella, China), DMEM medium (L110KJ, Yuanpei, China), Fetal bovine serum(AC03L055, life-ilab, China), GDF-6 (HY-P79333, MedChemexpress, USA), GDF-7 (92004ES10, Yisheng, China), CTGF (HY-P72154, MedChemexpress Biotechnology, USA), GoldenstarTM RT6 cDNA Synthesis Kit Ver.2 (TSK302M, Tsingke, China), qPCR Mix (SYBR Green I) kit and PCR primer (TSE002, Tsingke, China), RIPA lysis buffer (Beyotime, China), TNC (A1927, abclonal, China), MKX (ab236400, abcam, UK), SCX (ab58655, abcam, UK), GAPDH (A19056, abcam, UK), Hes1 (ab71559, abcam, UK), Hras (ab32417, abcam, UK), GAPDH Rabbit mAb (A19056, abclonal, China), Lentiviral Packaging Kit (41102ES10, Yisheng, China), Fetal bovine serum (26050070, Thermo, USA), trypsin (LP0042, Thermo, USA), PBS (10010001, Thermo, USA).

Techniques: Expressing, Immunofluorescence, Control

Journal: Molecular Medicine Reports

Article Title: Molecular responses of radiation-induced liver damage in rats

doi: 10.3892/mmr.2014.3051

Figure Lengend Snippet: Immunohistochemical staining of liver sections 3 days and 1, 2, 4, 8 and 12 weeks after 20 Gy irradiation. Brown color denotes positivity. The perivenous area was the major area of positive staining. TGF-β1, Smad3, CTGF and NF-κB p65 staining was positive between 3 days and 12 weeks after irradiation. Smad4 and Smad7 staining was positive between 3 days and 1 week after irradiation. TNF-α staining was positive between 1 and 4 weeks after irradiation. (Magnification, ×200). Gy, grays; TGF-β1, transforming growth factor-β1; CTGF, connective tissue growth factor; TNFα, tumor necrosis factor-α, NF, nuclear factor.

Article Snippet: The primary antibodies used were as follows: Rabbit monoclonal antibodies against transforming growth factor-β1 (TGF-β1; 1:250), nuclear factor (NF)-κB65 (1:100), mothers against decapentaplegic homolog 4 (Smad4) (1:40), Smad3 (1:250), Smad7 (1:30), tumor necrosis factor-α (TNF-α; 1:200) and connective tissue growth factor (CTGF; 1:200), all purchased from Boster Co., Ltd.

Techniques: Immunohistochemical staining, Staining, Irradiation

Journal: Molecular Medicine Reports

Article Title: Molecular responses of radiation-induced liver damage in rats

doi: 10.3892/mmr.2014.3051

Figure Lengend Snippet: Western blot analysis (ratio of the molecules investigated, vs. GAPDH). Similar to the results obtained using the reverse transcription quantitative polymerase chain reaction, the protein expression levels were as follows: NF-κB p65 was increased between 3 days and 12 weeks after irradiation. CTGF and Smad3 were significantly increased between 2 and 12 weeks after irradiation. TGF-β1 was significantly increased between 1 and 12 weeks after irradiation and TNF-α was significantly increased between 3 days and 4 weeks after irradiation. Smad4 was significantly increased between 3 days and 1 week after irradiation. Smad7 was significantly increased 3 days after irradiation and reduced significantly 1 and 2 weeks after irradiation, however, it remained higher than that in the control. From 4 weeks post-irradiation, the protein expression of Smad7 returned to the control level. * P<0.05, ** P<0.001 compared with the control group (0 Gy). Gy, grays; CTGF, connective tissue growth factor; TGF-β1, transforming growth factor-β1; TNFα, tumor necrosis factor-α; Smad, mothers against decapentaplegic; NF, nuclear factor.

Article Snippet: The primary antibodies used were as follows: Rabbit monoclonal antibodies against transforming growth factor-β1 (TGF-β1; 1:250), nuclear factor (NF)-κB65 (1:100), mothers against decapentaplegic homolog 4 (Smad4) (1:40), Smad3 (1:250), Smad7 (1:30), tumor necrosis factor-α (TNF-α; 1:200) and connective tissue growth factor (CTGF; 1:200), all purchased from Boster Co., Ltd.

Techniques: Western Blot, Reverse Transcription, Real-time Polymerase Chain Reaction, Expressing, Irradiation, Control

Journal: Molecular Medicine Reports

Article Title: Molecular responses of radiation-induced liver damage in rats

doi: 10.3892/mmr.2014.3051

Figure Lengend Snippet: mRNA expression levels were determined using reverse transcription quantitative polymerase chain reaction (ratio of the molecules investigated, vs. GAPDH). The mRNA expression of NF-κB p65 was upregulated between 3 days and 12 weeks after irradiation. The mRNA expression levels were as follows: CTGF and Smad3 were significantly upregulated between 2 and 12 weeks after irradiation. TGF-β1 was significantly upregulated between 1 and 12 weeks after irradiation. TNF-α was significantly upregulated between 3 days and 4 weeks after irradiation. Smad4 was significantly upregulated 3 days and 1 week after irradiation. Smad7 was significantly upregulated 3 days after irradiation and reduced significantly 1–2 weeks after irradiation, however, the expression levels remained higher than that in the control. From 4 weeks post-irradiation, the mRNA expression of Smad7 returned to the control level. * P<0.05, ** P<0.001, compared with the control group (0 Gy). CTGF, connective tissue growth factor; TGF-β1, transforming growth factor-β1; TNFα, tumor necrosis factor-α, Smad, mothers against decapentaplegic; NF, nuclear factor.

Article Snippet: The primary antibodies used were as follows: Rabbit monoclonal antibodies against transforming growth factor-β1 (TGF-β1; 1:250), nuclear factor (NF)-κB65 (1:100), mothers against decapentaplegic homolog 4 (Smad4) (1:40), Smad3 (1:250), Smad7 (1:30), tumor necrosis factor-α (TNF-α; 1:200) and connective tissue growth factor (CTGF; 1:200), all purchased from Boster Co., Ltd.

Techniques: Expressing, Reverse Transcription, Real-time Polymerase Chain Reaction, Irradiation, Control