human il11 elisa kits  (CUSAg Inc)

Journal: Cell Death & Disease

Article Title: MCM7 promotes liver fibrosis by transcriptionally regulating IL11 via the SHCBP1-RACGAP1-STAT3 axis

doi: 10.1038/s41419-025-07937-x

Figure Lengend Snippet: A , B Volcano plots of differentially expressed genes in MCM7-knockdown ( A ) or SHCBP1-knockdown ( B ) cells treated with IL-1β. Upregulated genes are highlighted in red, and downregulated genes in blue. C , D Venn diagrams illustrating the overlap of upregulated ( C ) and downregulated ( D ) differentially expressed genes between MCM7-knockdown and SHCBP1-knockdown cells upon IL-1β treatment. E , F Pathway analysis of the upregulated overlapping genes ( E ) and downregulated overlapping genes ( F ). G Table listing significantly differentially expressed genes from the cytokine-cytokine receptor interaction network. H Heatmap depicting the downregulated expression of genes in both MCM7-knockdown and SHCBP1-knockdown cells upon IL-1β treatment. I Genome browser tracks of RNA-Seq signals at the IL11 gene in PLKO.1, MCM7-knockdown, and SHCBP1-knockdown cells after IL-1β treatment.

Article Snippet: The levels of IL11 in human and mouse serum samples were determined using the human IL11 ELISA Kit (CHE0010-096, 4A Biotech, Beijing, China) and the mouse IL11 ELISA Kit (CME0007-096, 4A Biotech, Beijing, China), respectively.

Techniques: Knockdown, Expressing, RNA Sequencing

Journal: Cell Death & Disease

Article Title: MCM7 promotes liver fibrosis by transcriptionally regulating IL11 via the SHCBP1-RACGAP1-STAT3 axis

doi: 10.1038/s41419-025-07937-x

Figure Lengend Snippet: A Identification of SHCBP1-interacting proteins by mass spectrometry, with the top ten interacting partners shown. B The peptide map from the marked protein band identifies RACGAP1 as a SHCBP1-interacting protein. C Co-IP analysis of the endogenous interaction between SHCBP1 and RACGAP1 in HepG2 cells. D Co-IP showing the interaction between RACGAP1 and STAT3 in HepG2 cells transfected with shCtrl or shSHCBP1. E Immunofluorescence (IF) showing the colocalization of STAT3 (red), RACGAP1 (green), and DAPI (nucleus, blue) in HepG2 cells transfected with shCtrl or shSHCBP1 (scale bar: 5 μm). The graph presents quantitative analysis of RACGAP1/STAT3 colocalization. F , G Co-IP analysis showing the interaction between SHCBP1 and RACGAP1 ( F ) or between RACGAP1 and STAT3 ( G ) in HepG2 cells transfected with shCtrl or shMCM7. H IF showing the colocalization of STAT3 (red), RACGAP1 (green), and DAPI (nucleus, blue) in HepG2 cells transfected with shCtrl or shMCM7 (scale bar: 5 μm). The graph presents quantitative analysis of RACGAP1/STAT3 colocalization. I , J Western blot analysis of phosphorylated STAT3 (p-STAT3) in HepG2 cells with MCM7 knockdown and co-transfected with pHAGE-RACGAP1 ( I ) or pHAGE-SHCBP1 ( J ). K , M Western blot analysis of p-STAT3 in liver tissues from S. japonicum -infected mice treated with AAV-shMcm7 ( K ) or AAV-shShcbp1 ( M ). Graphs show the relative protein expression levels. L , N Correlation between p-STAT3 levels and MCM7 expression ( L ) or SHCBP1 expression ( N ) in liver tissues from S. japonicum -infected mice ( n = 12). O Relative luciferase activity of the IL11 promoter in HepG2 cells transfected with pHAGE or pHAGE-MCM7 with or without C188-9. P The schematic diagram shows the IL11 promoter region indicating STAT3 binding sites, and ChIP-qPCR analysis of STAT3, MCM7, and SHCBP1 binding to the IL11 promoter was conducted in HepG2 cells. Q Relative luciferase activity of the IL11 promoter in HepG2 cells transfected with pHAGE or pHAGE-MCM7, with or without mutations in the STAT3 binding site (−1348/−1337). Data are presented as mean ± SD of three independent experiments. Statistical analyses were performed using unpaired Student’s t -test, one-way ANOVA. * P < 0.05; *** P < 0.001; **** P < 0.0001; ns not significant.

Article Snippet: The levels of IL11 in human and mouse serum samples were determined using the human IL11 ELISA Kit (CHE0010-096, 4A Biotech, Beijing, China) and the mouse IL11 ELISA Kit (CME0007-096, 4A Biotech, Beijing, China), respectively.

Techniques: Mass Spectrometry, Co-Immunoprecipitation Assay, Transfection, Immunofluorescence, Western Blot, Knockdown, Infection, Expressing, Luciferase, Activity Assay, Binding Assay, ChIP-qPCR

Journal: Cell Death & Disease

Article Title: MCM7 promotes liver fibrosis by transcriptionally regulating IL11 via the SHCBP1-RACGAP1-STAT3 axis

doi: 10.1038/s41419-025-07937-x

Figure Lengend Snippet: A Experimental design schematic: HSCs isolated from uninfected mice were exposed to conditioned medium from primary hepatocytes of S. japonicum -infected mice treated with AAV-EGFP or AAV-MCM7, with or without IL11 neutralizing antibody (top). Relative mRNA expression levels of Col1a1 and α-Sma in HSCs were analyzed by qRT-PCR (bottom). B Experimental design schematic: LX-2 cells were exposed to conditioned medium from HepG2 cells transfected with pHAGE or pHAGE-MCM7, with or without IL11 neutralizing antibody (top). Relative mRNA expression levels of COL1A1 and α-SMA in LX-2 cells were analyzed by qRT-PCR (bottom). C Experimental design schematic: mice were infected with S. japonicum and intravenously injected with either AAV-MCM7 or AAV-EGFP on day 10 post-infection. Four weeks before sacrifice, mice were intraperitoneally treated rhIL11 every two days. Liver samples were collected at 8 weeks post-infection for analysis. D H&E staining (areas positive for liver fibrosis are delineated by black dashed lines), Masson’s trichrome staining, COL1A1 staining, and α-SMA staining (all scale bars: 100 μm) of liver sections from the indicated groups (AAV-EGFP + PBS + uninfected, AAV-EGFP + PBS + infected, AAV-MCM7 + PBS + infected, AAV-MCM7 + rhIL11 + infected). Graphs show the quantified positive areas for each stain, determined using ImageJ software from multiple randomly selected fields across distinct tissue sections. E Hydroxyproline content in liver tissues was determined. F The size of the granuloma area in S. japonicum -infected mice (AAV-EGFP + PBS + infected, AAV-MCM7 + PBS + infected, AAV-MCM7 + rhIL11 + infected) was measured and calculated. G , H qRT-PCR ( G ) was used to assess the expression levels of Col1a1 , α-Sma , Timp1 , and Mmp2 , while Western blot ( H ) analysis focused on COL1A1 and α-SMA in liver tissues from the indicated groups, with the graph displaying protein levels. Data are presented as mean ± SD of 3–6 mice per group and are representative of three independent experiments. Statistical analyses were performed using unpaired Student’s t -test or one-way ANOVA. * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001.

Article Snippet: The levels of IL11 in human and mouse serum samples were determined using the human IL11 ELISA Kit (CHE0010-096, 4A Biotech, Beijing, China) and the mouse IL11 ELISA Kit (CME0007-096, 4A Biotech, Beijing, China), respectively.

Techniques: Isolation, Infection, Expressing, Quantitative RT-PCR, Transfection, Injection, Staining, Software, Western Blot

Journal: Cancer Research

Article Title: BCA101 Is a Tumor-Targeted Bifunctional Fusion Antibody That Simultaneously Inhibits EGFR and TGFβ Signaling to Durably Suppress Tumor Growth

doi: 10.1158/0008-5472.can-21-4425

Figure Lengend Snippet: Figure 1. BCA101 retains EGFR and TGFb binding activity in fusion format. A, Computer-generated working model of BCA101 (front view). BCA101 is composed of anti-human EGFR mAb (cetuximab) and TGFbreceptor RIIECD fused to the C terminus of the LC of cetuximab IgGvia a flexible (G4S)3linker. B, A 90 rotation of BCA101 structure showing (Gly4Ser)3 linker connected to TGFbRII-ECD (side view). C, BCA101 reducing and nonreducing SDS-PAGE. Cetuximab and BCA101 were analyzed by SDS- PAGE under both reducing (lane 1 and 2, respectively) and nonreducing (lane 4 and 5, respectively) conditions. Lanes 3 and 6 are protein markers along with the expected molecular weight scaleon either side. D, SPR-based BCA101 bindingto itscognate targets, EGFR and TGFb. rh EGFR or TGFbwas immobilizedon CM5 chips. BCA101 or controls were run at a specified flow rate for a set of association and dissociation times to determine Kd values (nmol/L). E, FaDu cells were incubated with serial dilutions of BCA101 or cetuximab, followed by a fluorochrome-conjugated anti-human IgG secondary antibody. BCA101 and cetuximab binding to cells was analyzed by flow cytometer and MFI was plotted against concentration. EGFR and TGFb binding ELISA. F and G, Plates were coated with either EGFR-Fc for EGFR binding (F) or TGFb for TGFbRII-Fc binding (G) and probed with various concentrations of BCA101, followed by an HRP-conjugated secondary antibody specific to human IgG. Color was developed by adding TMB substrate and plates were read using a plate reader. H, BCA101 bifunctional ELISA. Plates were coated with EGFR-Fc, probed with various concentrations of BCA101, followed by TGFb spiking, biotinylated anti-TGFb antibody, and SA-HRP. Plates were developed with substrate (TMB) and read in a plate reader. Cetuximab was used as a negative control. Anti-VEGF mAb was used as an IgG control for E–G. Cetuximab was used as a negative control in bifunctional ELISA. For D, F, and G, three independent experiments were performed, and representative data are presented as mean SD of triplicate. E, Only one experiment was performed. For H, six independent experiments were performed, and representative data are presented as mean SD of triplicate.

Article Snippet: After 120 hours, cell culture supernatant was collected from the assay plate and proceeded with IL11 ELISA as per manufacturer’s instructions using human IL11 Quantikine ELISA Kit (R&D Systems Inc.).

Techniques: Binding Assay, Activity Assay, Generated, SDS Page, Molecular Weight, Incubation, Cytometry, Concentration Assay, Enzyme-linked Immunosorbent Assay, Negative Control, Control

Journal: Cancer Research

Article Title: BCA101 Is a Tumor-Targeted Bifunctional Fusion Antibody That Simultaneously Inhibits EGFR and TGFβ Signaling to Durably Suppress Tumor Growth

doi: 10.1158/0008-5472.can-21-4425

Figure Lengend Snippet: Figure 5. BCA101 preferentially localizes to tumor and neutralizes TGFb. A and B, Biodistribution of BCA101 compared with cetuximab and TGFbRII-Fc. BCA101, cetuximab, and TGFbRII-Fc were labeled with Alexa Flour 647 to the similar degree of labeling and administered to FaDu xenograft-bearing mice (n ¼ 1, 10 mg/kg, i.p.). Whole animal dorsal imaging (A) was performed at defined time points using In-vivo Xtream imaging station (Bruker) and net intensity from tumor-defined ROI (B) obtained at each time point was plotted against time as ln of photons/second/mm2. C–E, TGFb neutralization in vivo. C, BALB/c mice (n ¼ 3) were injected with BCA101 at 5 mg/kg intravenously and serum samples were collected at defined time points. BCA101 and TGFb levels in serum were evaluated by ELISA. Data presented as mean SD of 3 mice at each time point. Dotted line, TGFb levels (150 pg/mL) in placebo-treated mice. A431 xenograft-bearing mice were treated (intraperitoneally, twice a week, seven doses) with BCA101, TGFbRII-Fc, or cetuximab (10 mg/kg; n ¼ minimum 9) for 19 days and tumors and serum were collected. Tumor lysates (D) and serum (E) were evaluated for TGFb levels by ELISA. Data are plotted as mean SD. Statistical analysis for D and E was performed using one-way ANOVA, nonparametric, Kruskal–Wallis test followed by Dunn multiple comparison test where mean rank of each column was compared with every other column. Significance is indicated by , P ≤0.05; , P ≤0.01; , P ≤0.001; , P ≤0.0001.

Article Snippet: After 120 hours, cell culture supernatant was collected from the assay plate and proceeded with IL11 ELISA as per manufacturer’s instructions using human IL11 Quantikine ELISA Kit (R&D Systems Inc.).

Techniques: Labeling, Imaging, In Vivo, Neutralization, Injection, Enzyme-linked Immunosorbent Assay, Comparison

Journal: Cancer Research

Article Title: BCA101 Is a Tumor-Targeted Bifunctional Fusion Antibody That Simultaneously Inhibits EGFR and TGFβ Signaling to Durably Suppress Tumor Growth

doi: 10.1158/0008-5472.can-21-4425

Figure Lengend Snippet: Figure 7. BCA101 in combination with immune checkpoint therapeutics. A and B, BCA101 and immune checkpoint inhibitor combination studies in vitro. In A and B, human PBMCs were stimulated with soluble anti-CD3 (2 ng/mL). A, Exogenous TGFb added at 10 ng/mL and drugs were added to a final concentration of 56 nmol/L. IFNg release was measured in supernatant after about 72 hours using ELISA. Absorbance values are plotted as mean SD from four independent experiments in triplicates. B, Activated human PBMCs were cocultured with HCT116 cells in presence of TGFb spiked at 1 ng/mL. Drugs were added to a final concentration of 6.667 nmol/L. IFNg level was measured in the supernatantafter about 72 hours and fold increasewas calculatedover hIgG for the test groups. Data were plotted as mean SD of six independent experiments each in triplicate. (Continued on the following page.)

Article Snippet: After 120 hours, cell culture supernatant was collected from the assay plate and proceeded with IL11 ELISA as per manufacturer’s instructions using human IL11 Quantikine ELISA Kit (R&D Systems Inc.).

Techniques: In Vitro, Concentration Assay, Enzyme-linked Immunosorbent Assay