Journal: Biomedicines

Article Title: Podocalyxin, Isthmin-1, and Pentraxin-3 Immunoreactivities as Emerging Immunohistochemical Markers of Fibrosis in Chronic Hepatitis B

doi: 10.3390/biomedicines13122958

Figure Lengend Snippet: The immunoreactivities of PODXL, PTX3 and ISM-1 in the low fibrosis, high fibrosis, and control groups. (Arrows indicate immunoreactivities).

Article Snippet: After washing the tissues three times for 5 min each with PBS, 5 min of Ultra V Block (TA–125-UB, Lab Vision Corporation) solution was applied to prevent background staining, followed by incubation with PTX3, PODXL, and ISM-1 primary antibodies diluted at a ratio of 1/200 (PTX3 Polyclonal Antibody, PA5-36156, Thermo Fisher Scientific, Invitrogen, Waltham, MA, USA/PODXL, 39-3800, Thermo Fisher Scientific Inc./ISM1 Polyclonal Antibody, E-AB-18133, Elabscience Biotechnology, Houston, TX, USA) for 60 min at room temperature in a humidified environment.

Techniques: Control

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 Radiographic comparison of hip joints between healthy controls and patients with ONFH. B ELISA assay of serum PTX3 levels between healthy controls and patients with ONFH. (n = 53, 32 control subjects and 21 case subjects, statistical analysis: Mann–Whitney U test.) C HE (n = 5 technical replicates) and IHC staining (n = 3 technical replicates) and quantitative analysis of femoral head tissue sections in patients with traumatic hip dislocation and femoral head necrosis. Scale bar: 100 μm. Statistical analysis: two-tailed Student’s t test. D qPCR analysis of Ptx3 in MC3T3-E1 treated with or without Dex and quantification, normalized to Gapdh (internal control). (n = 3 independent experiments) Statistical analysis: two-tailed Student’s t test. E WB analysis of PTX3 protein levels in MC3T3-E1 treated with or without Dex relative to β-Actin and quantification. (n = 3 independent experiments) Statistical analysis: two-tailed Student’s t test. F Immunofluorescence staining of PTX3 in MC3T3-E1 treated with or without Dex and quantification of relative fluorescence intensity. (n = 3 independent experiments) Statistical analysis: two-tailed Student’s t test. Error bars: standard deviation, SD. Red arrows: empty lacunae; White arrows: positive cells. Control: Standard osteogenic induction medium (OIM); Dex: Standard OIM co-cultured with dexamethasone (10 μM). 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 C57BL/6 Ptx3 -knockout mice were purchased from Cyagen Biosciences and the C57BL/6 wild-type mice were acquired from Beijing Vital River Laboratory Animal Technology Co., Ltd in Beijing, China.

Techniques: Comparison, Enzyme-linked Immunosorbent Assay, Control, MANN-WHITNEY, Immunohistochemistry, Two Tailed Test, Immunofluorescence, Staining, Fluorescence, Standard Deviation, Cell Culture

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 Flowchart of ONFH model on Ptx3 -KO and wild-type mice. B The appearance of femoral heads in both groups after ONFH modeling. C HE staining and Goldner trichrome staining of femoral heads in both groups and quantitative analysis of the results of HE staining (n = 5 technical replicates) and Goldner trichrome staining (n = 3 technical replicates). D IHC staining of ALP, OCN and COL1 in both groups and quantitative analysis of immunohistochemical positive expression in each group of mice. (n = 3 technical replicates). E SEM of femoral heads in both groups. F Micro-CT of femoral heads in both groups and related quantification (n = 5 technical replicates). Ptx3 +/+ group: wild-type mice of ONFH model, n = 5; Ptx3 −/− group: Ptx3 -knockout mice of ONFH model, n = 5. Statistical analysis: two-tailed Student’s t test. Error bars: standard deviation, SD. Red arrows: empty lacunae; Black arrows: trabecular bone disruption; 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 C57BL/6 Ptx3 -knockout mice were purchased from Cyagen Biosciences and the C57BL/6 wild-type mice were acquired from Beijing Vital River Laboratory Animal Technology Co., Ltd in Beijing, China.

Techniques: Staining, Immunohistochemistry, Immunohistochemical staining, Expressing, Micro-CT, Knock-Out, Two Tailed Test, Standard Deviation, 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: A The appearance of femoral heads in control, GIONFH model (Dex), and ATF3 inducer 1-treated groups. B HE staining of femoral heads in three groups and quantitative analysis of the results of HE staining (n = 5 technical replicates). C IHC staining of ALP, OCN and COL1 in three groups and quantitative analysis of immunohistochemical positive expression in each group of mice. (n = 3 technical replicates). D The appearance of femoral heads in ATF3 inducer 1-treated WT and Ptx3 -KO groups. E HE staining of femoral heads in both groups and quantitative analysis of the results of HE staining (n = 5 technical replicates). F IHC staining of ALP, OCN and COL1 in both groups and quantitative analysis of immunohistochemical positive expression in each group of mice. (n = 3 technical replicates). Statistical analysis: Dunnett’s post-hoc tests. Error bars: standard deviation, SD. Control: vehicle-treated, n = 5; Dex: GIONH model, n = 5; Dex+ATF3/ Ptx3 +/+ +ATF3: GIONFH model with 40 mg/kg ATF3 inducer 1 administration twice a week, n = 5; Ptx3 −/− +ATF3: Ptx3 -knockout mice with GIONFH modeling and ATF3 inducer 1, 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.

Article Snippet: The C57BL/6 Ptx3 -knockout mice were purchased from Cyagen Biosciences and the C57BL/6 wild-type mice were acquired from Beijing Vital River Laboratory Animal Technology Co., Ltd in Beijing, China.

Techniques: Control, Staining, Immunohistochemistry, Immunohistochemical staining, Expressing, Standard Deviation, Knock-Out, 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: 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 C57BL/6 Ptx3 -knockout mice were purchased from Cyagen Biosciences and the C57BL/6 wild-type mice were acquired from Beijing Vital River Laboratory Animal Technology Co., Ltd in Beijing, China.

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 C57BL/6 Ptx3 -knockout mice were purchased from Cyagen Biosciences and the C57BL/6 wild-type mice were acquired from Beijing Vital River Laboratory Animal Technology Co., Ltd in Beijing, China.

Techniques: