Journal: Gut and Liver

Article Title: Expression of Fibroblast Growth Factor 21 and β-Klotho Regulates Hepatic Fibrosis through the Nuclear Factor-κB and c-Jun N-Terminal Kinase Pathways

doi: 10.5009/gnl17443

Figure Lengend Snippet: Analysis of serum FGF19 and FGF21 levels in patients with viral or alcoholic hepatitis. Serum levels of FGF19 (A) and FGF21 (B) were determined by enzyme-linked immunosorbent assay. G1 (F0–F1, n=10), G2 (F2–F3, n=10), G3 (F4A–F4C, n=15). FGF, fibroblast growth factor. *p<0.01.

Article Snippet: The lysates were subjected to Western blotting analysis under reducing conditions using previously validated human β-Klotho antibodies (R & D Systems), anti-phospho-Erk1/2, total Erk1/2, phospho-AKT, total AKT, phospho-JNK, and total JNK antibodies (Cell Signaling Technology, Danvers, MA, USA), phospho-IκBα, total IκBα, proliferating cell nuclear antigen (PCNA), and GAPDH antibodies (Santa Cruz Biotechnology, Santa Cruz, CA, USA), and FGF21 antibody (Boster Biological Technology, Pleasanton, CA, USA).

Techniques: Enzyme-linked Immunosorbent Assay

Journal: Gut and Liver

Article Title: Expression of Fibroblast Growth Factor 21 and β-Klotho Regulates Hepatic Fibrosis through the Nuclear Factor-κB and c-Jun N-Terminal Kinase Pathways

doi: 10.5009/gnl17443

Figure Lengend Snippet: Analysis of FGF19, FGF21, and β-Klotho expression and inflammatory markers in liver tissue. Expression levels of mRNA for FGF19 (A), FGF21 (B), β-Klotho (C), IL-1β (D), IL-6 (E), and TNF-α (F) were determined by real-time polymerase chain reaction. G1 (F0–F1, n=10), G2 (F2–F3, n=10), G3 (F4A–F4C, n=15). FGF, fibroblast growth factor; IL, interleukin; TNF, tumor necrosis factor. *p<0.05, † p<0.01.

Article Snippet: The lysates were subjected to Western blotting analysis under reducing conditions using previously validated human β-Klotho antibodies (R & D Systems), anti-phospho-Erk1/2, total Erk1/2, phospho-AKT, total AKT, phospho-JNK, and total JNK antibodies (Cell Signaling Technology, Danvers, MA, USA), phospho-IκBα, total IκBα, proliferating cell nuclear antigen (PCNA), and GAPDH antibodies (Santa Cruz Biotechnology, Santa Cruz, CA, USA), and FGF21 antibody (Boster Biological Technology, Pleasanton, CA, USA).

Techniques: Expressing, Real-time Polymerase Chain Reaction

Journal: Gut and Liver

Article Title: Expression of Fibroblast Growth Factor 21 and β-Klotho Regulates Hepatic Fibrosis through the Nuclear Factor-κB and c-Jun N-Terminal Kinase Pathways

doi: 10.5009/gnl17443

Figure Lengend Snippet: Correlation between the Fibrosis Stage and mRNA Expression of FGF19/21, β-Klotho, and Inflammatory Markers

Article Snippet: The lysates were subjected to Western blotting analysis under reducing conditions using previously validated human β-Klotho antibodies (R & D Systems), anti-phospho-Erk1/2, total Erk1/2, phospho-AKT, total AKT, phospho-JNK, and total JNK antibodies (Cell Signaling Technology, Danvers, MA, USA), phospho-IκBα, total IκBα, proliferating cell nuclear antigen (PCNA), and GAPDH antibodies (Santa Cruz Biotechnology, Santa Cruz, CA, USA), and FGF21 antibody (Boster Biological Technology, Pleasanton, CA, USA).

Techniques: Expressing

Journal: Gut and Liver

Article Title: Expression of Fibroblast Growth Factor 21 and β-Klotho Regulates Hepatic Fibrosis through the Nuclear Factor-κB and c-Jun N-Terminal Kinase Pathways

doi: 10.5009/gnl17443

Figure Lengend Snippet: Effects of IL-1β on β-Klotho and FGF21 in Huh-7 cells. Huh-7 cells were incubated with IL-1β for 6 hours. β-Klotho expression levels (A and G), protein kinase B (B), extracelluar signal-regulated kinases (C), JNK (D), IκBα (E), and FGF21 (F and G) were determined by immunoblotting. IL, interleukin; FGF, fibroblast growth factor; JNK, c-Jun N-terminal kinase.

Article Snippet: The lysates were subjected to Western blotting analysis under reducing conditions using previously validated human β-Klotho antibodies (R & D Systems), anti-phospho-Erk1/2, total Erk1/2, phospho-AKT, total AKT, phospho-JNK, and total JNK antibodies (Cell Signaling Technology, Danvers, MA, USA), phospho-IκBα, total IκBα, proliferating cell nuclear antigen (PCNA), and GAPDH antibodies (Santa Cruz Biotechnology, Santa Cruz, CA, USA), and FGF21 antibody (Boster Biological Technology, Pleasanton, CA, USA).

Techniques: Incubation, Expressing, Western Blot

Journal: Gut and Liver

Article Title: Expression of Fibroblast Growth Factor 21 and β-Klotho Regulates Hepatic Fibrosis through the Nuclear Factor-κB and c-Jun N-Terminal Kinase Pathways

doi: 10.5009/gnl17443

Figure Lengend Snippet: Signaling pathways that inhibit β-Klotho and induce FGF21 by IL-1β in Huh-7 cells. Huh-7 cells were pretreated with an NF-κB inhibitor (Bay11-7082, A and E), JNK inhibitor (SP600125, B and F), protein kinase B inhibitor (LY294002, C and G), or extracelluar signal-regulated kinases inhibitor (PD98059, D and H) for 20 minutes and then treated with 10 ng/mL of IL-1β for 6 hours to detect β-Klotho and FGF21. (I) Huh-7 cells were pretreated with 1 μM NF-κB inhibitor Bay11-7082, after which β-Klotho and FGF21 levels were determined by immunoblotting. (J) Huh-7 cells were pretreated with 0.5 μM JNK inhibitor SP600125, after which β-Klotho and FGF21 levels were determined by immunoblotting. (K) β-Klotho and FGF21 levels were assessed 6 hours after pretreatment with 1 μM NF-κB inhibitor Bay11-7082 in Huh-7 cells. (L) β-Klotho and FGF21 levels were assessed 6 hours after pretreatment with 0.5 μM JNK inhibitor SP600125 in Huh-7 cells. FGF, fibroblast growth factor; IL, interleukin; NF-κB, nuclear factor-κB; JNK, c-Jun N-terminal kinase; GAPDH, glyceraldehyde-3-phosphate dehydrogenase.

Article Snippet: The lysates were subjected to Western blotting analysis under reducing conditions using previously validated human β-Klotho antibodies (R & D Systems), anti-phospho-Erk1/2, total Erk1/2, phospho-AKT, total AKT, phospho-JNK, and total JNK antibodies (Cell Signaling Technology, Danvers, MA, USA), phospho-IκBα, total IκBα, proliferating cell nuclear antigen (PCNA), and GAPDH antibodies (Santa Cruz Biotechnology, Santa Cruz, CA, USA), and FGF21 antibody (Boster Biological Technology, Pleasanton, CA, USA).

Techniques: Protein-Protein interactions, Western Blot

Journal: Gut and Liver

Article Title: Expression of Fibroblast Growth Factor 21 and β-Klotho Regulates Hepatic Fibrosis through the Nuclear Factor-κB and c-Jun N-Terminal Kinase Pathways

doi: 10.5009/gnl17443

Figure Lengend Snippet: FGF21 inhibits IL-1β-induced growth retardation of hepatocytes. Huh-7 cells were incubated with 10 ng/mL of IL-1β (A), 500 ng/mL of FGF21 (B), or IL-1β+FGF21 (C), after which the expression of β-Klotho and PCNA was determined. (D) IL-1β-treated Huh-7 cells were incubated with or without FGF21 in a dose-dependent manner. Cell viability was measured by an methylthiazolyldiphenyl-tetrazolium bromide (MTT) assay. IL, interleukin; FGF, fibroblast growth factor; PCNA, proliferating cell nuclear antigen; GAPDH, glyceraldehyde-3-phosphate dehydrogenase.

Article Snippet: The lysates were subjected to Western blotting analysis under reducing conditions using previously validated human β-Klotho antibodies (R & D Systems), anti-phospho-Erk1/2, total Erk1/2, phospho-AKT, total AKT, phospho-JNK, and total JNK antibodies (Cell Signaling Technology, Danvers, MA, USA), phospho-IκBα, total IκBα, proliferating cell nuclear antigen (PCNA), and GAPDH antibodies (Santa Cruz Biotechnology, Santa Cruz, CA, USA), and FGF21 antibody (Boster Biological Technology, Pleasanton, CA, USA).

Techniques: Incubation, Expressing, MTT Assay

Journal: Communications Biology

Article Title: Pentraxin 3 ameliorates glucocorticoid-induced osteonecrosis of the femoral head via TLR4/NF-κB/FGF21 signaling axis

doi: 10.1038/s42003-025-09282-3

Figure Lengend Snippet: A RNA sequencing pre-processing flowchart. B Signaling pathways up-regulated in rPTX3-treated MC3T3-E1. C Volcano map of differential genes between rPTX3 treatment and Dex groups. D Heat map of differential genes of the cytokines and growth factors between rPTX3 treatment and Dex groups based on RNA sequencing. E qPCR analysis of Fgf21, Gdf15, Hgf, Ereg, Tgfb2 , and Fgf10 in Dex stimulated MC3T3-E1 treated with or without rPTX3 and quantification, normalized to Gapdh (internal control). F WB analysis and relative quantification of FGF21 in Dex stimulated MC3T3-E1 treated with or without rPTX3. Actin was used as an internal control. G qPCR analysis of Fgf21 in shFGF21 infected MC3T3-E1. H WB analysis and quantification of FGF21 in shFGF21 infected MC3T3-E1. Actin was used as an internal control. I WB analysis and quantification of osteogenesis markers (ALP, Runx2 and COL1 at day 7, OCN and OPN at day 14) and apoptosis markers (Bcl-2 and Bax at 24 h) in five groups of MC3T3-E1. Actin was used as an internal control. J Cell death/live analysis in five groups and quantification. K Flow cytometry analysis in five groups and quantification. (Apoptotic cells: Q2 + Q3) L , M ALP staining and ARS staining in five groups and quantification. N , O Immunofluorescence staining and quantitative analysis of relative FI of osteogenesis markers OCN (day 14), Runx2 (day 7) in five groups. Control: MC3T3 cells cultured with standard OIM, n = 3; shFGF21: shFGF21-MC3T3 cells cultured with standard OIM, n = 3; Dex: MC3T3 cells cultured with standard OIM and dexamethasone (10 μM), n = 3; shFGF21+Dex: shFGF21-MC3T3 cells cultured with standard OIM and dexamethasone (10 μM). Dex+rFGF21: MC3T3 cells cultured with standard OIM co-cultured with dexamethasone (10 μM) and rFGF21 (200 ng/mL), n = 3. Statistical analysis: Dunnett’s post-hoc tests (n = 3 independent experiments). Error bars: standard deviation, SD. The images provided in all figures represent typical examples from each experimental group. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: The sections were then incubated overnight at 4 °C with primary antibodies targeting PTX3 (1:200 dilution, 13797-1-AP, Proteintech), osteocalcin (OCN, 1:200 dilution, DF12303, Affinity), alkaline phosphatase (ALP, 1:200 dilution, DF6225, Affinity), Collagen I (COL1, 1:500 dilution, A24112, ABclonal), phosphorylated p65 (p-p65,1:200 dilution, AF2006, Affinity), phosphorylated IκB (p-IκB, 1:1000 dilution, 82349-1-RR, Proteintech) and FGF21 (1:200 dilution, TC52978S, Abmart).

Techniques: RNA Sequencing, Protein-Protein interactions, Control, Quantitative Proteomics, Infection, Flow Cytometry, Staining, Immunofluorescence, Cell Culture, Standard Deviation

Journal: Communications Biology

Article Title: Pentraxin 3 ameliorates glucocorticoid-induced osteonecrosis of the femoral head via TLR4/NF-κB/FGF21 signaling axis

doi: 10.1038/s42003-025-09282-3

Figure Lengend Snippet: A , B FGF2 and FGF21 exhibit certain structural similarities at the protein level. C Co-Immunoprecipitation analysis of FGF21 and PTX3 interaction. D KEGG enrichment analysis and heatmap visualization of TLR/NF-κB pathway-associated differentially expressed genes. E WB analysis and quantification of FGF21 and TLR4/NF-κB pathway-associated markers TLR4, MyD88, p-p65/p65 and p-IκB/IκB in five groups of MC3T3-E1. Actin was used as an internal control. F Cell death/live analysis in five groups. G Flow cytometry analysis in five groups. (Apoptotic cells: Q2 + Q3). H , I ALP staining and ARS staining in five groups. J , K Immunofluorescence staining of osteogenesis markers OCN (Day 14), Runx2 (Day 7) in five groups. L The appearance of femoral heads in five groups. M HE staining of femoral heads in five groups. N IHC staining of ALP, OCN, and COL1 in five groups. (n = 3 independent experiments) Statistical analysis: Dunnett’s post-hoc tests. Error bars: standard deviation, SD. Control: Standard OIM, n = 3/ vehicle-treated, n = 5; Dex: Standard OIM co-cultured with dexamethasone (10 μM), n = 3/ glucocorticoid-induced osteonecrosis model, n = 5; Dex+rPTX3: Standard OIM co-cultured with dexamethasone (10 uM) and rPTX3 (200 ng/mL), n = 3/ model + 200 ng/g recombinant PTX3, n = 5; Dex+rPTX3+TAK-242: Standard OIM co-cultured with dexamethasone (10 uM), rPTX3 (200 ng/mL) and TAK-242 (50 nM), n = 3/ model + rPTX3 + TAK-242, n = 5; Dex+rPTX3 + SN50: Standard OIM co-cultured with dexamethasone (10 μM), rPTX3 (200 ng/mL) and SN50 (50 μg/mL), n = 3/ model + rPTX3 + SN50, n = 5. Red arrows: empty lacunae; Black arrows: trabecular bone disruption; Star symbol: necrotic bone marrow; White arrows: positive cells. The images provided in all figures represent typical examples from each experimental group. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: The sections were then incubated overnight at 4 °C with primary antibodies targeting PTX3 (1:200 dilution, 13797-1-AP, Proteintech), osteocalcin (OCN, 1:200 dilution, DF12303, Affinity), alkaline phosphatase (ALP, 1:200 dilution, DF6225, Affinity), Collagen I (COL1, 1:500 dilution, A24112, ABclonal), phosphorylated p65 (p-p65,1:200 dilution, AF2006, Affinity), phosphorylated IκB (p-IκB, 1:1000 dilution, 82349-1-RR, Proteintech) and FGF21 (1:200 dilution, TC52978S, Abmart).

Techniques: Immunoprecipitation, Control, Flow Cytometry, Staining, Immunofluorescence, Immunohistochemistry, Standard Deviation, Cell Culture, Recombinant, Disruption

Journal: Communications Biology

Article Title: Pentraxin 3 ameliorates glucocorticoid-induced osteonecrosis of the femoral head via TLR4/NF-κB/FGF21 signaling axis

doi: 10.1038/s42003-025-09282-3

Figure Lengend Snippet: Schematic diagram of PTX3-mediated amelioration of glucocorticoid-induced osteonecrosis in the femoral head via TLR4/NF-κB/FGF21 signaling axis.

Article Snippet: The sections were then incubated overnight at 4 °C with primary antibodies targeting PTX3 (1:200 dilution, 13797-1-AP, Proteintech), osteocalcin (OCN, 1:200 dilution, DF12303, Affinity), alkaline phosphatase (ALP, 1:200 dilution, DF6225, Affinity), Collagen I (COL1, 1:500 dilution, A24112, ABclonal), phosphorylated p65 (p-p65,1:200 dilution, AF2006, Affinity), phosphorylated IκB (p-IκB, 1:1000 dilution, 82349-1-RR, Proteintech) and FGF21 (1:200 dilution, TC52978S, Abmart).

Techniques: