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human tinagl1 elisa kit  (Boster Bio)


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    Boster Bio human tinagl1 elisa kit
    Figure 1. <t>TINAGL1</t> is increased in HCV-infected patients, hepatocytes and in the liver of mice and patients with fibrosis. (A) Venn diagram analysis of genes in HCV-infected Huh7.5 cells at the indicated time points (Fold change ≥ 2). (B-C) RNA levels quantified by qRT-PCR (B) and proteins detected by Western blot (C) in Huh7.5 cells infected with HCV for over three months (n = 3). (D-E) RNA (D) and protein (E) levels in Huh7.5 cells infected with HCV (MOI = 0.1) for 24, 48, and 72 hours (n = 3). (F-G) RNA (F) and protein (G) levels in Huh7.5 cells infected with HCV (MOI = 0.01, 0.1, and 1) for 72 hours (n = 3). (H) TINAGL1 quantified by ELISA in the culture medium of HCV-infected Huh7.5 cells (n = 3). (I) TINAGL1 mRNA in liver biopsies from Gene Expression Omnibus database (GSE38226). (J) TINAGL1 measured by Western blot in mouse livers with fibrosis. (K-L) Statistical summary of TINAGL1 expression in a human liver tissue array from patients with MASH (n = 29), fibrosis (n = 92), cirrhosis (n = 30), and normal donors (n = 10). Data were expressed as mean ± standard deviation. P values were calculated by an unpaired two-tailed Student’s t-test (B, H, I, and L) or one-way ANOVA (D, F, and K) using Tukey’s multiple comparisons test. α-SMA, alpha-smooth muscle actin; CCl4, carbon tetrachloride; DEN, diethylnitrosamine; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; HCV, hepatitis C virus; TINAGL1, tubulointerstitial nephritis antigen-like 1.
    Human Tinagl1 Elisa Kit, supplied by Boster Bio, used in various techniques. Bioz Stars score: 94/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/human tinagl1 elisa kit/product/Boster Bio
    Average 94 stars, based on 2 article reviews
    human tinagl1 elisa kit - by Bioz Stars, 2026-02
    94/100 stars

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    1) Product Images from "Tubulointerstitial nephritis antigen-like 1 promotes the progression of liver fibrosis after HCV eradication with direct-acting antivirals."

    Article Title: Tubulointerstitial nephritis antigen-like 1 promotes the progression of liver fibrosis after HCV eradication with direct-acting antivirals.

    Journal: International journal of biological sciences

    doi: 10.7150/ijbs.103305

    Figure 1. TINAGL1 is increased in HCV-infected patients, hepatocytes and in the liver of mice and patients with fibrosis. (A) Venn diagram analysis of genes in HCV-infected Huh7.5 cells at the indicated time points (Fold change ≥ 2). (B-C) RNA levels quantified by qRT-PCR (B) and proteins detected by Western blot (C) in Huh7.5 cells infected with HCV for over three months (n = 3). (D-E) RNA (D) and protein (E) levels in Huh7.5 cells infected with HCV (MOI = 0.1) for 24, 48, and 72 hours (n = 3). (F-G) RNA (F) and protein (G) levels in Huh7.5 cells infected with HCV (MOI = 0.01, 0.1, and 1) for 72 hours (n = 3). (H) TINAGL1 quantified by ELISA in the culture medium of HCV-infected Huh7.5 cells (n = 3). (I) TINAGL1 mRNA in liver biopsies from Gene Expression Omnibus database (GSE38226). (J) TINAGL1 measured by Western blot in mouse livers with fibrosis. (K-L) Statistical summary of TINAGL1 expression in a human liver tissue array from patients with MASH (n = 29), fibrosis (n = 92), cirrhosis (n = 30), and normal donors (n = 10). Data were expressed as mean ± standard deviation. P values were calculated by an unpaired two-tailed Student’s t-test (B, H, I, and L) or one-way ANOVA (D, F, and K) using Tukey’s multiple comparisons test. α-SMA, alpha-smooth muscle actin; CCl4, carbon tetrachloride; DEN, diethylnitrosamine; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; HCV, hepatitis C virus; TINAGL1, tubulointerstitial nephritis antigen-like 1.
    Figure Legend Snippet: Figure 1. TINAGL1 is increased in HCV-infected patients, hepatocytes and in the liver of mice and patients with fibrosis. (A) Venn diagram analysis of genes in HCV-infected Huh7.5 cells at the indicated time points (Fold change ≥ 2). (B-C) RNA levels quantified by qRT-PCR (B) and proteins detected by Western blot (C) in Huh7.5 cells infected with HCV for over three months (n = 3). (D-E) RNA (D) and protein (E) levels in Huh7.5 cells infected with HCV (MOI = 0.1) for 24, 48, and 72 hours (n = 3). (F-G) RNA (F) and protein (G) levels in Huh7.5 cells infected with HCV (MOI = 0.01, 0.1, and 1) for 72 hours (n = 3). (H) TINAGL1 quantified by ELISA in the culture medium of HCV-infected Huh7.5 cells (n = 3). (I) TINAGL1 mRNA in liver biopsies from Gene Expression Omnibus database (GSE38226). (J) TINAGL1 measured by Western blot in mouse livers with fibrosis. (K-L) Statistical summary of TINAGL1 expression in a human liver tissue array from patients with MASH (n = 29), fibrosis (n = 92), cirrhosis (n = 30), and normal donors (n = 10). Data were expressed as mean ± standard deviation. P values were calculated by an unpaired two-tailed Student’s t-test (B, H, I, and L) or one-way ANOVA (D, F, and K) using Tukey’s multiple comparisons test. α-SMA, alpha-smooth muscle actin; CCl4, carbon tetrachloride; DEN, diethylnitrosamine; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; HCV, hepatitis C virus; TINAGL1, tubulointerstitial nephritis antigen-like 1.

    Techniques Used: Infection, Quantitative RT-PCR, Western Blot, Enzyme-linked Immunosorbent Assay, Gene Expression, Expressing, Standard Deviation, Two Tailed Test, Virus

    Figure 2. TINAGL1 expression remains higher in hepatocytes after HCV elimination by high-efficiency treatment with DAAs. (A) Schematic diagram (left) for the establishment of HCV-eradicated Huh7.5 cells and HCV RNA (right) in the cells (n = 3). (B) TINAGL1 quantified by ELISA in the culture medium of HCV-eradicated Huh7.5 cells (n = 3). (C-D) mRNA (C) and protein (D) of TINAGL1 in HCV-eradicated Huh7.5 cells (n = 3). (E) Protein levels in HCV-eradicated Huh7.5 cells after re-infection with HCV for 72 hours (n = 3). Data were expressed as mean ± standard deviation. P values were calculated by one-way ANOVA (B, C) using Tukey’s multiple comparisons test. DAAs, direct-acting antivirals.
    Figure Legend Snippet: Figure 2. TINAGL1 expression remains higher in hepatocytes after HCV elimination by high-efficiency treatment with DAAs. (A) Schematic diagram (left) for the establishment of HCV-eradicated Huh7.5 cells and HCV RNA (right) in the cells (n = 3). (B) TINAGL1 quantified by ELISA in the culture medium of HCV-eradicated Huh7.5 cells (n = 3). (C-D) mRNA (C) and protein (D) of TINAGL1 in HCV-eradicated Huh7.5 cells (n = 3). (E) Protein levels in HCV-eradicated Huh7.5 cells after re-infection with HCV for 72 hours (n = 3). Data were expressed as mean ± standard deviation. P values were calculated by one-way ANOVA (B, C) using Tukey’s multiple comparisons test. DAAs, direct-acting antivirals.

    Techniques Used: Expressing, Enzyme-linked Immunosorbent Assay, Infection, Standard Deviation

    Figure 3. TINAGL1 promotes the activation of hepatic stellate cells in vitro. (A) Schematic diagram of the transwell insert mono- and co-culture models. (B-C) mRNA levels in LX-2 cells co-cultured with HCV-infected Huh7.5 cells (B) or their CM (C) (n = 5). (D-E) mRNA levels in LX-2 cells co-cultured with Huh7.5 cells transfected with the TINAGL1 plasmid (D) or their CM (E) (n = 5). (F) mRNA levels in LX-2 cells treated with rhTINAGL1 for 48 hours (n = 3). Data were expressed as mean ± standard deviation. P values were calculated by an unpaired two-tailed Student’s t-test (B-E) or one-way ANOVA (F) using Tukey’s multiple comparisons test. ACTA2 (α-SMA), alpha-smooth muscle actin; CM conditioned medium; COL1A1, collagen type I alpha 1; rhTINAGL1, recombinant human tubulointerstitial nephritis antigen-like 1; TIMP1, tissue inhibitor of matrix metalloproteinase 1.
    Figure Legend Snippet: Figure 3. TINAGL1 promotes the activation of hepatic stellate cells in vitro. (A) Schematic diagram of the transwell insert mono- and co-culture models. (B-C) mRNA levels in LX-2 cells co-cultured with HCV-infected Huh7.5 cells (B) or their CM (C) (n = 5). (D-E) mRNA levels in LX-2 cells co-cultured with Huh7.5 cells transfected with the TINAGL1 plasmid (D) or their CM (E) (n = 5). (F) mRNA levels in LX-2 cells treated with rhTINAGL1 for 48 hours (n = 3). Data were expressed as mean ± standard deviation. P values were calculated by an unpaired two-tailed Student’s t-test (B-E) or one-way ANOVA (F) using Tukey’s multiple comparisons test. ACTA2 (α-SMA), alpha-smooth muscle actin; CM conditioned medium; COL1A1, collagen type I alpha 1; rhTINAGL1, recombinant human tubulointerstitial nephritis antigen-like 1; TIMP1, tissue inhibitor of matrix metalloproteinase 1.

    Techniques Used: Activation Assay, In Vitro, Co-Culture Assay, Cell Culture, Infection, Transfection, Plasmid Preparation, Standard Deviation, Two Tailed Test, Recombinant

    Figure 4. TINAGL1 activates HSCs by stabilizing PDGF-BB. (A) Scatter Chart of fibrosis-associated cytokines in the culture medium of Huh7.5 cells transfected with the TINAGL1 plasmid detected by a cytokine microarray assay. (B) PDGF-BB concentration quantified by ELISA in cell lysate and culture medium and by Western Blot in cell lysate of Huh7.5 cells transfected with the TINAGL1 plasmid (n = 3). (C) PDGFB mRNA level in Huh7.5 cells transfected with the TINAGL1 plasmid (n = 4). (D) Protein levels in Huh7.5 cells transfected with the TINAGL1-Flag plasmid and treated with cycloheximide (CHX), the intensity of protein was scanned by Image J (n =3). (E) Binding mode between human TINAGL1 and PDGF-BB. (F) Binding affinity between rhTINAGL1 (C-6 × His) and rhPDGF-BB proteins detected by SPR method. (G) Interaction of TINAGL1 and PDGF-BB in HEK293T cells detected by co-immunoprecipitation. (H) Co-localization of TINAGL1 (red) and PDGF-BB (green) in HEK293T cells (Scale bar: 5 μm). Nuclear (blue). Statistical analysis of co-localization of TINAGL1 and PDGF-BB fluorescence intensities was performed using ImageJ software. Data were expressed as mean ± standard deviation. P values were calculated by an unpaired two-tailed Student’s t-test (B-D). ACTB, beta-actin; CHX, cycloheximide; PDGF-BB, platelet-derived growth factor BB; rhPDGF-BB, recombinant human platelet-derived growth factor BB; rhTINAGL1, recombinant human tubulointerstitial nephritis antigen-like 1; SPR, surface plasmon resonance.
    Figure Legend Snippet: Figure 4. TINAGL1 activates HSCs by stabilizing PDGF-BB. (A) Scatter Chart of fibrosis-associated cytokines in the culture medium of Huh7.5 cells transfected with the TINAGL1 plasmid detected by a cytokine microarray assay. (B) PDGF-BB concentration quantified by ELISA in cell lysate and culture medium and by Western Blot in cell lysate of Huh7.5 cells transfected with the TINAGL1 plasmid (n = 3). (C) PDGFB mRNA level in Huh7.5 cells transfected with the TINAGL1 plasmid (n = 4). (D) Protein levels in Huh7.5 cells transfected with the TINAGL1-Flag plasmid and treated with cycloheximide (CHX), the intensity of protein was scanned by Image J (n =3). (E) Binding mode between human TINAGL1 and PDGF-BB. (F) Binding affinity between rhTINAGL1 (C-6 × His) and rhPDGF-BB proteins detected by SPR method. (G) Interaction of TINAGL1 and PDGF-BB in HEK293T cells detected by co-immunoprecipitation. (H) Co-localization of TINAGL1 (red) and PDGF-BB (green) in HEK293T cells (Scale bar: 5 μm). Nuclear (blue). Statistical analysis of co-localization of TINAGL1 and PDGF-BB fluorescence intensities was performed using ImageJ software. Data were expressed as mean ± standard deviation. P values were calculated by an unpaired two-tailed Student’s t-test (B-D). ACTB, beta-actin; CHX, cycloheximide; PDGF-BB, platelet-derived growth factor BB; rhPDGF-BB, recombinant human platelet-derived growth factor BB; rhTINAGL1, recombinant human tubulointerstitial nephritis antigen-like 1; SPR, surface plasmon resonance.

    Techniques Used: Transfection, Plasmid Preparation, Microarray, Concentration Assay, Enzyme-linked Immunosorbent Assay, Western Blot, Binding Assay, Immunoprecipitation, Fluorescence, Software, Standard Deviation, Two Tailed Test, Derivative Assay, Recombinant, SPR Assay

    Figure 5. TINAGL1 activates HSCs by PDGF-BB/PDGFRβ pathway. (A) PDGFRB mRNA levels in LX-2 cells treated with rhTINAGL1 for 48 hours (n = 3). (B-C) Protein levels of α-SMA and PDGFRβ in LX-2 cell and TINAGL1 in cell medium after co-culture with Huh7.5 cells transfected with TINAGL1 plasmid, siRNA (B) or neutralizing antibody (C) against PDGFRβ, the intensity of protein was scanned by Image J (n =3). Data were expressed as mean ± standard deviation. P values were calculated by one-way ANOVA (A-C) using Tukey’s multiple comparisons test. α-SMA, alpha-smooth muscle actin; PDGFRβ, platelet-derived growth factor receptor beta; rhTINAGL1, recombinant human tubulointerstitial nephritis antigen-like 1.
    Figure Legend Snippet: Figure 5. TINAGL1 activates HSCs by PDGF-BB/PDGFRβ pathway. (A) PDGFRB mRNA levels in LX-2 cells treated with rhTINAGL1 for 48 hours (n = 3). (B-C) Protein levels of α-SMA and PDGFRβ in LX-2 cell and TINAGL1 in cell medium after co-culture with Huh7.5 cells transfected with TINAGL1 plasmid, siRNA (B) or neutralizing antibody (C) against PDGFRβ, the intensity of protein was scanned by Image J (n =3). Data were expressed as mean ± standard deviation. P values were calculated by one-way ANOVA (A-C) using Tukey’s multiple comparisons test. α-SMA, alpha-smooth muscle actin; PDGFRβ, platelet-derived growth factor receptor beta; rhTINAGL1, recombinant human tubulointerstitial nephritis antigen-like 1.

    Techniques Used: Co-Culture Assay, Transfection, Plasmid Preparation, Standard Deviation, Derivative Assay, Recombinant

    Figure 6. Liver-specific overexpression of TINAGL1 initiates and exacerbates liver fibrosis in mice. (A) Schematic overview of the experimental setup for (B-C). (B) H&E staining (Scale bar: 100 μm), Masson staining (Scale bar: 100 μm), and Sirius red staining (Scale bar: 100 μm) of mouse livers. Quantification of Masson and Sirius red
    Figure Legend Snippet: Figure 6. Liver-specific overexpression of TINAGL1 initiates and exacerbates liver fibrosis in mice. (A) Schematic overview of the experimental setup for (B-C). (B) H&E staining (Scale bar: 100 μm), Masson staining (Scale bar: 100 μm), and Sirius red staining (Scale bar: 100 μm) of mouse livers. Quantification of Masson and Sirius red

    Techniques Used: Over Expression, Staining

    Figure 8. Liver-specific knockdown of TINAGL1 prevents the progression of liver fibrosis in mice induced by CCl4. (A) Schematic overview of the experimental setup. (B) mRNA level of Tinagl1 in mouse livers (n = 5). (C) Body weight (n = 5). (D) H&E staining (Scale bar: 100 μm), Masson staining (Scale bar: 100 μm), and Sirius Red staining (Scale bar: 100 μm) of mouse livers. Quantification of Masson and Sirius red staining with Image J. (E) Liver / body weight ratio (n = 5). (F) Hydroxyproline content in mouse livers (n = 5). (G) Serum AST and ALT in mice (n = 5). (H) mRNA and protein levels in mouse livers (n = 5). (I) Serum PDGF-BB level quantified by ELISA (n = 5). Data were expressed as mean ± standard deviation. P values were calculated by one-way ANOVA (B, D-I) using Tukey’s multiple comparisons test or two-way ANOVA (C) using Bonferroni’s multiple comparisons test. Acta2 (α-SMA), alpha-smooth muscle actin; AST, aspartate aminotransferase; ALT alanine aminotransferase; COL1A1, collagen type I alpha 1; PDGFRβ, platelet-derived growth factor receptor beta; TIMP1, tissue inhibitor of matrix metalloproteinase 1; Il1b, interleukin-1 beta; Il6, interleukin 6; Tnfa, tumor necrosis factor alpha.
    Figure Legend Snippet: Figure 8. Liver-specific knockdown of TINAGL1 prevents the progression of liver fibrosis in mice induced by CCl4. (A) Schematic overview of the experimental setup. (B) mRNA level of Tinagl1 in mouse livers (n = 5). (C) Body weight (n = 5). (D) H&E staining (Scale bar: 100 μm), Masson staining (Scale bar: 100 μm), and Sirius Red staining (Scale bar: 100 μm) of mouse livers. Quantification of Masson and Sirius red staining with Image J. (E) Liver / body weight ratio (n = 5). (F) Hydroxyproline content in mouse livers (n = 5). (G) Serum AST and ALT in mice (n = 5). (H) mRNA and protein levels in mouse livers (n = 5). (I) Serum PDGF-BB level quantified by ELISA (n = 5). Data were expressed as mean ± standard deviation. P values were calculated by one-way ANOVA (B, D-I) using Tukey’s multiple comparisons test or two-way ANOVA (C) using Bonferroni’s multiple comparisons test. Acta2 (α-SMA), alpha-smooth muscle actin; AST, aspartate aminotransferase; ALT alanine aminotransferase; COL1A1, collagen type I alpha 1; PDGFRβ, platelet-derived growth factor receptor beta; TIMP1, tissue inhibitor of matrix metalloproteinase 1; Il1b, interleukin-1 beta; Il6, interleukin 6; Tnfa, tumor necrosis factor alpha.

    Techniques Used: Knockdown, Staining, Enzyme-linked Immunosorbent Assay, Standard Deviation, Derivative Assay

    Figure 9. Liver-specific knockdown of TINAGL1 alleviates liver fibrosis in mice induced by CCl4. (A) Schematic overview of the experimental setup. (B) mRNA level of Tinagl1 in mouse livers (n = 5). (C) Body weight (n = 5). (D) H&E staining (Scale bar: 100 μm), Masson staining (Scale bar: 100 μm), and Sirius Red staining (Scale bar: 100 μm) of mouse livers. Quantification of Masson and Sirius red staining with Image J. (E) Liver / body weight ratio (n = 5). (F) Hydroxyproline content in mouse livers (n = 5). (G) Serum ALT and AST in mice (n = 5). (H) mRNA and protein levels in mouse livers (n = 5). (I) Serum PDGF-BB level quantified by ELISA (n = 5). Data were expressed as mean ± standard deviation. P values were calculated by one-way ANOVA (B, D-I) using Tukey’s multiple comparisons test or two-way ANOVA (C) using Bonferroni’s multiple comparisons test. Acta2 (α-SMA), alpha-smooth muscle actin; AST, aspartate aminotransferase; ALT alanine aminotransferase; COL1A1, collagen type I alpha 1; PDGFRβ, platelet-derived growth factor receptor beta; TIMP1, tissue inhibitor of matrix metalloproteinase 1; Il1b, interleukin-1 beta; Il6, interleukin 6; Tnfa, tumor necrosis factor alpha.
    Figure Legend Snippet: Figure 9. Liver-specific knockdown of TINAGL1 alleviates liver fibrosis in mice induced by CCl4. (A) Schematic overview of the experimental setup. (B) mRNA level of Tinagl1 in mouse livers (n = 5). (C) Body weight (n = 5). (D) H&E staining (Scale bar: 100 μm), Masson staining (Scale bar: 100 μm), and Sirius Red staining (Scale bar: 100 μm) of mouse livers. Quantification of Masson and Sirius red staining with Image J. (E) Liver / body weight ratio (n = 5). (F) Hydroxyproline content in mouse livers (n = 5). (G) Serum ALT and AST in mice (n = 5). (H) mRNA and protein levels in mouse livers (n = 5). (I) Serum PDGF-BB level quantified by ELISA (n = 5). Data were expressed as mean ± standard deviation. P values were calculated by one-way ANOVA (B, D-I) using Tukey’s multiple comparisons test or two-way ANOVA (C) using Bonferroni’s multiple comparisons test. Acta2 (α-SMA), alpha-smooth muscle actin; AST, aspartate aminotransferase; ALT alanine aminotransferase; COL1A1, collagen type I alpha 1; PDGFRβ, platelet-derived growth factor receptor beta; TIMP1, tissue inhibitor of matrix metalloproteinase 1; Il1b, interleukin-1 beta; Il6, interleukin 6; Tnfa, tumor necrosis factor alpha.

    Techniques Used: Knockdown, Staining, Enzyme-linked Immunosorbent Assay, Standard Deviation, Derivative Assay



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    human tinagl1 elisa kit  (Boster Bio)
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    Boster Bio human tinagl1 elisa kit
    Figure 1. <t>TINAGL1</t> is increased in HCV-infected patients, hepatocytes and in the liver of mice and patients with fibrosis. (A) Venn diagram analysis of genes in HCV-infected Huh7.5 cells at the indicated time points (Fold change ≥ 2). (B-C) RNA levels quantified by qRT-PCR (B) and proteins detected by Western blot (C) in Huh7.5 cells infected with HCV for over three months (n = 3). (D-E) RNA (D) and protein (E) levels in Huh7.5 cells infected with HCV (MOI = 0.1) for 24, 48, and 72 hours (n = 3). (F-G) RNA (F) and protein (G) levels in Huh7.5 cells infected with HCV (MOI = 0.01, 0.1, and 1) for 72 hours (n = 3). (H) TINAGL1 quantified by ELISA in the culture medium of HCV-infected Huh7.5 cells (n = 3). (I) TINAGL1 mRNA in liver biopsies from Gene Expression Omnibus database (GSE38226). (J) TINAGL1 measured by Western blot in mouse livers with fibrosis. (K-L) Statistical summary of TINAGL1 expression in a human liver tissue array from patients with MASH (n = 29), fibrosis (n = 92), cirrhosis (n = 30), and normal donors (n = 10). Data were expressed as mean ± standard deviation. P values were calculated by an unpaired two-tailed Student’s t-test (B, H, I, and L) or one-way ANOVA (D, F, and K) using Tukey’s multiple comparisons test. α-SMA, alpha-smooth muscle actin; CCl4, carbon tetrachloride; DEN, diethylnitrosamine; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; HCV, hepatitis C virus; TINAGL1, tubulointerstitial nephritis antigen-like 1.
    Human Tinagl1 Elisa Kit, supplied by Boster Bio, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/human tinagl1 elisa kit/product/Boster Bio
    Average 94 stars, based on 1 article reviews
    human tinagl1 elisa kit - by Bioz Stars, 2026-02
    94/100 stars
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    Figure 1. TINAGL1 is increased in HCV-infected patients, hepatocytes and in the liver of mice and patients with fibrosis. (A) Venn diagram analysis of genes in HCV-infected Huh7.5 cells at the indicated time points (Fold change ≥ 2). (B-C) RNA levels quantified by qRT-PCR (B) and proteins detected by Western blot (C) in Huh7.5 cells infected with HCV for over three months (n = 3). (D-E) RNA (D) and protein (E) levels in Huh7.5 cells infected with HCV (MOI = 0.1) for 24, 48, and 72 hours (n = 3). (F-G) RNA (F) and protein (G) levels in Huh7.5 cells infected with HCV (MOI = 0.01, 0.1, and 1) for 72 hours (n = 3). (H) TINAGL1 quantified by ELISA in the culture medium of HCV-infected Huh7.5 cells (n = 3). (I) TINAGL1 mRNA in liver biopsies from Gene Expression Omnibus database (GSE38226). (J) TINAGL1 measured by Western blot in mouse livers with fibrosis. (K-L) Statistical summary of TINAGL1 expression in a human liver tissue array from patients with MASH (n = 29), fibrosis (n = 92), cirrhosis (n = 30), and normal donors (n = 10). Data were expressed as mean ± standard deviation. P values were calculated by an unpaired two-tailed Student’s t-test (B, H, I, and L) or one-way ANOVA (D, F, and K) using Tukey’s multiple comparisons test. α-SMA, alpha-smooth muscle actin; CCl4, carbon tetrachloride; DEN, diethylnitrosamine; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; HCV, hepatitis C virus; TINAGL1, tubulointerstitial nephritis antigen-like 1.

    Journal: International journal of biological sciences

    Article Title: Tubulointerstitial nephritis antigen-like 1 promotes the progression of liver fibrosis after HCV eradication with direct-acting antivirals.

    doi: 10.7150/ijbs.103305

    Figure Lengend Snippet: Figure 1. TINAGL1 is increased in HCV-infected patients, hepatocytes and in the liver of mice and patients with fibrosis. (A) Venn diagram analysis of genes in HCV-infected Huh7.5 cells at the indicated time points (Fold change ≥ 2). (B-C) RNA levels quantified by qRT-PCR (B) and proteins detected by Western blot (C) in Huh7.5 cells infected with HCV for over three months (n = 3). (D-E) RNA (D) and protein (E) levels in Huh7.5 cells infected with HCV (MOI = 0.1) for 24, 48, and 72 hours (n = 3). (F-G) RNA (F) and protein (G) levels in Huh7.5 cells infected with HCV (MOI = 0.01, 0.1, and 1) for 72 hours (n = 3). (H) TINAGL1 quantified by ELISA in the culture medium of HCV-infected Huh7.5 cells (n = 3). (I) TINAGL1 mRNA in liver biopsies from Gene Expression Omnibus database (GSE38226). (J) TINAGL1 measured by Western blot in mouse livers with fibrosis. (K-L) Statistical summary of TINAGL1 expression in a human liver tissue array from patients with MASH (n = 29), fibrosis (n = 92), cirrhosis (n = 30), and normal donors (n = 10). Data were expressed as mean ± standard deviation. P values were calculated by an unpaired two-tailed Student’s t-test (B, H, I, and L) or one-way ANOVA (D, F, and K) using Tukey’s multiple comparisons test. α-SMA, alpha-smooth muscle actin; CCl4, carbon tetrachloride; DEN, diethylnitrosamine; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; HCV, hepatitis C virus; TINAGL1, tubulointerstitial nephritis antigen-like 1.

    Article Snippet: The human TINAGL1 ELISA kit (Boster, EK1766) was used to examine TINAGL1 concentration in culture medium and in cell lysates of Huh7.5 cells.

    Techniques: Infection, Quantitative RT-PCR, Western Blot, Enzyme-linked Immunosorbent Assay, Gene Expression, Expressing, Standard Deviation, Two Tailed Test, Virus

    Figure 2. TINAGL1 expression remains higher in hepatocytes after HCV elimination by high-efficiency treatment with DAAs. (A) Schematic diagram (left) for the establishment of HCV-eradicated Huh7.5 cells and HCV RNA (right) in the cells (n = 3). (B) TINAGL1 quantified by ELISA in the culture medium of HCV-eradicated Huh7.5 cells (n = 3). (C-D) mRNA (C) and protein (D) of TINAGL1 in HCV-eradicated Huh7.5 cells (n = 3). (E) Protein levels in HCV-eradicated Huh7.5 cells after re-infection with HCV for 72 hours (n = 3). Data were expressed as mean ± standard deviation. P values were calculated by one-way ANOVA (B, C) using Tukey’s multiple comparisons test. DAAs, direct-acting antivirals.

    Journal: International journal of biological sciences

    Article Title: Tubulointerstitial nephritis antigen-like 1 promotes the progression of liver fibrosis after HCV eradication with direct-acting antivirals.

    doi: 10.7150/ijbs.103305

    Figure Lengend Snippet: Figure 2. TINAGL1 expression remains higher in hepatocytes after HCV elimination by high-efficiency treatment with DAAs. (A) Schematic diagram (left) for the establishment of HCV-eradicated Huh7.5 cells and HCV RNA (right) in the cells (n = 3). (B) TINAGL1 quantified by ELISA in the culture medium of HCV-eradicated Huh7.5 cells (n = 3). (C-D) mRNA (C) and protein (D) of TINAGL1 in HCV-eradicated Huh7.5 cells (n = 3). (E) Protein levels in HCV-eradicated Huh7.5 cells after re-infection with HCV for 72 hours (n = 3). Data were expressed as mean ± standard deviation. P values were calculated by one-way ANOVA (B, C) using Tukey’s multiple comparisons test. DAAs, direct-acting antivirals.

    Article Snippet: The human TINAGL1 ELISA kit (Boster, EK1766) was used to examine TINAGL1 concentration in culture medium and in cell lysates of Huh7.5 cells.

    Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Infection, Standard Deviation

    Figure 3. TINAGL1 promotes the activation of hepatic stellate cells in vitro. (A) Schematic diagram of the transwell insert mono- and co-culture models. (B-C) mRNA levels in LX-2 cells co-cultured with HCV-infected Huh7.5 cells (B) or their CM (C) (n = 5). (D-E) mRNA levels in LX-2 cells co-cultured with Huh7.5 cells transfected with the TINAGL1 plasmid (D) or their CM (E) (n = 5). (F) mRNA levels in LX-2 cells treated with rhTINAGL1 for 48 hours (n = 3). Data were expressed as mean ± standard deviation. P values were calculated by an unpaired two-tailed Student’s t-test (B-E) or one-way ANOVA (F) using Tukey’s multiple comparisons test. ACTA2 (α-SMA), alpha-smooth muscle actin; CM conditioned medium; COL1A1, collagen type I alpha 1; rhTINAGL1, recombinant human tubulointerstitial nephritis antigen-like 1; TIMP1, tissue inhibitor of matrix metalloproteinase 1.

    Journal: International journal of biological sciences

    Article Title: Tubulointerstitial nephritis antigen-like 1 promotes the progression of liver fibrosis after HCV eradication with direct-acting antivirals.

    doi: 10.7150/ijbs.103305

    Figure Lengend Snippet: Figure 3. TINAGL1 promotes the activation of hepatic stellate cells in vitro. (A) Schematic diagram of the transwell insert mono- and co-culture models. (B-C) mRNA levels in LX-2 cells co-cultured with HCV-infected Huh7.5 cells (B) or their CM (C) (n = 5). (D-E) mRNA levels in LX-2 cells co-cultured with Huh7.5 cells transfected with the TINAGL1 plasmid (D) or their CM (E) (n = 5). (F) mRNA levels in LX-2 cells treated with rhTINAGL1 for 48 hours (n = 3). Data were expressed as mean ± standard deviation. P values were calculated by an unpaired two-tailed Student’s t-test (B-E) or one-way ANOVA (F) using Tukey’s multiple comparisons test. ACTA2 (α-SMA), alpha-smooth muscle actin; CM conditioned medium; COL1A1, collagen type I alpha 1; rhTINAGL1, recombinant human tubulointerstitial nephritis antigen-like 1; TIMP1, tissue inhibitor of matrix metalloproteinase 1.

    Article Snippet: The human TINAGL1 ELISA kit (Boster, EK1766) was used to examine TINAGL1 concentration in culture medium and in cell lysates of Huh7.5 cells.

    Techniques: Activation Assay, In Vitro, Co-Culture Assay, Cell Culture, Infection, Transfection, Plasmid Preparation, Standard Deviation, Two Tailed Test, Recombinant

    Figure 4. TINAGL1 activates HSCs by stabilizing PDGF-BB. (A) Scatter Chart of fibrosis-associated cytokines in the culture medium of Huh7.5 cells transfected with the TINAGL1 plasmid detected by a cytokine microarray assay. (B) PDGF-BB concentration quantified by ELISA in cell lysate and culture medium and by Western Blot in cell lysate of Huh7.5 cells transfected with the TINAGL1 plasmid (n = 3). (C) PDGFB mRNA level in Huh7.5 cells transfected with the TINAGL1 plasmid (n = 4). (D) Protein levels in Huh7.5 cells transfected with the TINAGL1-Flag plasmid and treated with cycloheximide (CHX), the intensity of protein was scanned by Image J (n =3). (E) Binding mode between human TINAGL1 and PDGF-BB. (F) Binding affinity between rhTINAGL1 (C-6 × His) and rhPDGF-BB proteins detected by SPR method. (G) Interaction of TINAGL1 and PDGF-BB in HEK293T cells detected by co-immunoprecipitation. (H) Co-localization of TINAGL1 (red) and PDGF-BB (green) in HEK293T cells (Scale bar: 5 μm). Nuclear (blue). Statistical analysis of co-localization of TINAGL1 and PDGF-BB fluorescence intensities was performed using ImageJ software. Data were expressed as mean ± standard deviation. P values were calculated by an unpaired two-tailed Student’s t-test (B-D). ACTB, beta-actin; CHX, cycloheximide; PDGF-BB, platelet-derived growth factor BB; rhPDGF-BB, recombinant human platelet-derived growth factor BB; rhTINAGL1, recombinant human tubulointerstitial nephritis antigen-like 1; SPR, surface plasmon resonance.

    Journal: International journal of biological sciences

    Article Title: Tubulointerstitial nephritis antigen-like 1 promotes the progression of liver fibrosis after HCV eradication with direct-acting antivirals.

    doi: 10.7150/ijbs.103305

    Figure Lengend Snippet: Figure 4. TINAGL1 activates HSCs by stabilizing PDGF-BB. (A) Scatter Chart of fibrosis-associated cytokines in the culture medium of Huh7.5 cells transfected with the TINAGL1 plasmid detected by a cytokine microarray assay. (B) PDGF-BB concentration quantified by ELISA in cell lysate and culture medium and by Western Blot in cell lysate of Huh7.5 cells transfected with the TINAGL1 plasmid (n = 3). (C) PDGFB mRNA level in Huh7.5 cells transfected with the TINAGL1 plasmid (n = 4). (D) Protein levels in Huh7.5 cells transfected with the TINAGL1-Flag plasmid and treated with cycloheximide (CHX), the intensity of protein was scanned by Image J (n =3). (E) Binding mode between human TINAGL1 and PDGF-BB. (F) Binding affinity between rhTINAGL1 (C-6 × His) and rhPDGF-BB proteins detected by SPR method. (G) Interaction of TINAGL1 and PDGF-BB in HEK293T cells detected by co-immunoprecipitation. (H) Co-localization of TINAGL1 (red) and PDGF-BB (green) in HEK293T cells (Scale bar: 5 μm). Nuclear (blue). Statistical analysis of co-localization of TINAGL1 and PDGF-BB fluorescence intensities was performed using ImageJ software. Data were expressed as mean ± standard deviation. P values were calculated by an unpaired two-tailed Student’s t-test (B-D). ACTB, beta-actin; CHX, cycloheximide; PDGF-BB, platelet-derived growth factor BB; rhPDGF-BB, recombinant human platelet-derived growth factor BB; rhTINAGL1, recombinant human tubulointerstitial nephritis antigen-like 1; SPR, surface plasmon resonance.

    Article Snippet: The human TINAGL1 ELISA kit (Boster, EK1766) was used to examine TINAGL1 concentration in culture medium and in cell lysates of Huh7.5 cells.

    Techniques: Transfection, Plasmid Preparation, Microarray, Concentration Assay, Enzyme-linked Immunosorbent Assay, Western Blot, Binding Assay, Immunoprecipitation, Fluorescence, Software, Standard Deviation, Two Tailed Test, Derivative Assay, Recombinant, SPR Assay

    Figure 5. TINAGL1 activates HSCs by PDGF-BB/PDGFRβ pathway. (A) PDGFRB mRNA levels in LX-2 cells treated with rhTINAGL1 for 48 hours (n = 3). (B-C) Protein levels of α-SMA and PDGFRβ in LX-2 cell and TINAGL1 in cell medium after co-culture with Huh7.5 cells transfected with TINAGL1 plasmid, siRNA (B) or neutralizing antibody (C) against PDGFRβ, the intensity of protein was scanned by Image J (n =3). Data were expressed as mean ± standard deviation. P values were calculated by one-way ANOVA (A-C) using Tukey’s multiple comparisons test. α-SMA, alpha-smooth muscle actin; PDGFRβ, platelet-derived growth factor receptor beta; rhTINAGL1, recombinant human tubulointerstitial nephritis antigen-like 1.

    Journal: International journal of biological sciences

    Article Title: Tubulointerstitial nephritis antigen-like 1 promotes the progression of liver fibrosis after HCV eradication with direct-acting antivirals.

    doi: 10.7150/ijbs.103305

    Figure Lengend Snippet: Figure 5. TINAGL1 activates HSCs by PDGF-BB/PDGFRβ pathway. (A) PDGFRB mRNA levels in LX-2 cells treated with rhTINAGL1 for 48 hours (n = 3). (B-C) Protein levels of α-SMA and PDGFRβ in LX-2 cell and TINAGL1 in cell medium after co-culture with Huh7.5 cells transfected with TINAGL1 plasmid, siRNA (B) or neutralizing antibody (C) against PDGFRβ, the intensity of protein was scanned by Image J (n =3). Data were expressed as mean ± standard deviation. P values were calculated by one-way ANOVA (A-C) using Tukey’s multiple comparisons test. α-SMA, alpha-smooth muscle actin; PDGFRβ, platelet-derived growth factor receptor beta; rhTINAGL1, recombinant human tubulointerstitial nephritis antigen-like 1.

    Article Snippet: The human TINAGL1 ELISA kit (Boster, EK1766) was used to examine TINAGL1 concentration in culture medium and in cell lysates of Huh7.5 cells.

    Techniques: Co-Culture Assay, Transfection, Plasmid Preparation, Standard Deviation, Derivative Assay, Recombinant

    Figure 6. Liver-specific overexpression of TINAGL1 initiates and exacerbates liver fibrosis in mice. (A) Schematic overview of the experimental setup for (B-C). (B) H&E staining (Scale bar: 100 μm), Masson staining (Scale bar: 100 μm), and Sirius red staining (Scale bar: 100 μm) of mouse livers. Quantification of Masson and Sirius red

    Journal: International journal of biological sciences

    Article Title: Tubulointerstitial nephritis antigen-like 1 promotes the progression of liver fibrosis after HCV eradication with direct-acting antivirals.

    doi: 10.7150/ijbs.103305

    Figure Lengend Snippet: Figure 6. Liver-specific overexpression of TINAGL1 initiates and exacerbates liver fibrosis in mice. (A) Schematic overview of the experimental setup for (B-C). (B) H&E staining (Scale bar: 100 μm), Masson staining (Scale bar: 100 μm), and Sirius red staining (Scale bar: 100 μm) of mouse livers. Quantification of Masson and Sirius red

    Article Snippet: The human TINAGL1 ELISA kit (Boster, EK1766) was used to examine TINAGL1 concentration in culture medium and in cell lysates of Huh7.5 cells.

    Techniques: Over Expression, Staining

    Figure 8. Liver-specific knockdown of TINAGL1 prevents the progression of liver fibrosis in mice induced by CCl4. (A) Schematic overview of the experimental setup. (B) mRNA level of Tinagl1 in mouse livers (n = 5). (C) Body weight (n = 5). (D) H&E staining (Scale bar: 100 μm), Masson staining (Scale bar: 100 μm), and Sirius Red staining (Scale bar: 100 μm) of mouse livers. Quantification of Masson and Sirius red staining with Image J. (E) Liver / body weight ratio (n = 5). (F) Hydroxyproline content in mouse livers (n = 5). (G) Serum AST and ALT in mice (n = 5). (H) mRNA and protein levels in mouse livers (n = 5). (I) Serum PDGF-BB level quantified by ELISA (n = 5). Data were expressed as mean ± standard deviation. P values were calculated by one-way ANOVA (B, D-I) using Tukey’s multiple comparisons test or two-way ANOVA (C) using Bonferroni’s multiple comparisons test. Acta2 (α-SMA), alpha-smooth muscle actin; AST, aspartate aminotransferase; ALT alanine aminotransferase; COL1A1, collagen type I alpha 1; PDGFRβ, platelet-derived growth factor receptor beta; TIMP1, tissue inhibitor of matrix metalloproteinase 1; Il1b, interleukin-1 beta; Il6, interleukin 6; Tnfa, tumor necrosis factor alpha.

    Journal: International journal of biological sciences

    Article Title: Tubulointerstitial nephritis antigen-like 1 promotes the progression of liver fibrosis after HCV eradication with direct-acting antivirals.

    doi: 10.7150/ijbs.103305

    Figure Lengend Snippet: Figure 8. Liver-specific knockdown of TINAGL1 prevents the progression of liver fibrosis in mice induced by CCl4. (A) Schematic overview of the experimental setup. (B) mRNA level of Tinagl1 in mouse livers (n = 5). (C) Body weight (n = 5). (D) H&E staining (Scale bar: 100 μm), Masson staining (Scale bar: 100 μm), and Sirius Red staining (Scale bar: 100 μm) of mouse livers. Quantification of Masson and Sirius red staining with Image J. (E) Liver / body weight ratio (n = 5). (F) Hydroxyproline content in mouse livers (n = 5). (G) Serum AST and ALT in mice (n = 5). (H) mRNA and protein levels in mouse livers (n = 5). (I) Serum PDGF-BB level quantified by ELISA (n = 5). Data were expressed as mean ± standard deviation. P values were calculated by one-way ANOVA (B, D-I) using Tukey’s multiple comparisons test or two-way ANOVA (C) using Bonferroni’s multiple comparisons test. Acta2 (α-SMA), alpha-smooth muscle actin; AST, aspartate aminotransferase; ALT alanine aminotransferase; COL1A1, collagen type I alpha 1; PDGFRβ, platelet-derived growth factor receptor beta; TIMP1, tissue inhibitor of matrix metalloproteinase 1; Il1b, interleukin-1 beta; Il6, interleukin 6; Tnfa, tumor necrosis factor alpha.

    Article Snippet: The human TINAGL1 ELISA kit (Boster, EK1766) was used to examine TINAGL1 concentration in culture medium and in cell lysates of Huh7.5 cells.

    Techniques: Knockdown, Staining, Enzyme-linked Immunosorbent Assay, Standard Deviation, Derivative Assay

    Figure 9. Liver-specific knockdown of TINAGL1 alleviates liver fibrosis in mice induced by CCl4. (A) Schematic overview of the experimental setup. (B) mRNA level of Tinagl1 in mouse livers (n = 5). (C) Body weight (n = 5). (D) H&E staining (Scale bar: 100 μm), Masson staining (Scale bar: 100 μm), and Sirius Red staining (Scale bar: 100 μm) of mouse livers. Quantification of Masson and Sirius red staining with Image J. (E) Liver / body weight ratio (n = 5). (F) Hydroxyproline content in mouse livers (n = 5). (G) Serum ALT and AST in mice (n = 5). (H) mRNA and protein levels in mouse livers (n = 5). (I) Serum PDGF-BB level quantified by ELISA (n = 5). Data were expressed as mean ± standard deviation. P values were calculated by one-way ANOVA (B, D-I) using Tukey’s multiple comparisons test or two-way ANOVA (C) using Bonferroni’s multiple comparisons test. Acta2 (α-SMA), alpha-smooth muscle actin; AST, aspartate aminotransferase; ALT alanine aminotransferase; COL1A1, collagen type I alpha 1; PDGFRβ, platelet-derived growth factor receptor beta; TIMP1, tissue inhibitor of matrix metalloproteinase 1; Il1b, interleukin-1 beta; Il6, interleukin 6; Tnfa, tumor necrosis factor alpha.

    Journal: International journal of biological sciences

    Article Title: Tubulointerstitial nephritis antigen-like 1 promotes the progression of liver fibrosis after HCV eradication with direct-acting antivirals.

    doi: 10.7150/ijbs.103305

    Figure Lengend Snippet: Figure 9. Liver-specific knockdown of TINAGL1 alleviates liver fibrosis in mice induced by CCl4. (A) Schematic overview of the experimental setup. (B) mRNA level of Tinagl1 in mouse livers (n = 5). (C) Body weight (n = 5). (D) H&E staining (Scale bar: 100 μm), Masson staining (Scale bar: 100 μm), and Sirius Red staining (Scale bar: 100 μm) of mouse livers. Quantification of Masson and Sirius red staining with Image J. (E) Liver / body weight ratio (n = 5). (F) Hydroxyproline content in mouse livers (n = 5). (G) Serum ALT and AST in mice (n = 5). (H) mRNA and protein levels in mouse livers (n = 5). (I) Serum PDGF-BB level quantified by ELISA (n = 5). Data were expressed as mean ± standard deviation. P values were calculated by one-way ANOVA (B, D-I) using Tukey’s multiple comparisons test or two-way ANOVA (C) using Bonferroni’s multiple comparisons test. Acta2 (α-SMA), alpha-smooth muscle actin; AST, aspartate aminotransferase; ALT alanine aminotransferase; COL1A1, collagen type I alpha 1; PDGFRβ, platelet-derived growth factor receptor beta; TIMP1, tissue inhibitor of matrix metalloproteinase 1; Il1b, interleukin-1 beta; Il6, interleukin 6; Tnfa, tumor necrosis factor alpha.

    Article Snippet: The human TINAGL1 ELISA kit (Boster, EK1766) was used to examine TINAGL1 concentration in culture medium and in cell lysates of Huh7.5 cells.

    Techniques: Knockdown, Staining, Enzyme-linked Immunosorbent Assay, Standard Deviation, Derivative Assay