Anti-FGF2 Mouse Monoclonal Antibody [Clone ID: OTI3D9]

HEK293T cells were transfected with the pCMV6-ENTRY control (Left lane) or pCMV6-ENTRY BFGF (Right lane) cDNA for 48 hrs and lysed. Equivalent amounts of cell lysates (5 ug per lane) were separated by SDS-PAGE and immunoblotted with anti-BFGF.

Western blot analysis of extracts (10ug) from 4 different cell lines by using anti-BFGF monoclonal antibody at 1:200 dilution.

Immunohistochemical staining of paraffin-embedded Human pancreas tissue within the normal limits using anti-BFGF mouse monoclonal antibody. (Heat-induced epitope retrieval by 10mM citric buffer

Anti-BFGF mouse monoclonal antibody (M00121-2) immunofluorescent staining of COS7 cells transiently transfected by pCMV6-ENTRY BFGF.

Flow chart of the experimental design and verification of FGF2 deletion. a Diagram of experimental design. Bone marrow precursor cells were collected from the tibia and femur of WT and FGF2 KO mice aged 8–12 weeks. They were differentiated into BMDM for 7 days using 100ng/ml M-CSF. Macrophages in C57BL/6 male mice were depleted using clodronate liposomes. BMDM from WT and KO mice were injected via the tail vein into mice to reconstitute macrophages after 2 days. Mice were treated with Sham or CLP, and the samples were analyzed 24 h later. b Serum FGF2 protein levels were assessed by ELISA ( n = 3–5). c FGF2 gene expression levels were measured in the lungs, spleen, and liver using real-time PCR ( n = 3). d Western blot analysis of FGF2 protein levels in lung tissue extractions ( n = 3). e Immunofluorescence staining of FGF2 in BMDM from WT and FGF2 KO mice ( n = 3). f The levels of FGF2 gene expression were quantified in the lung tissue of mice subjected to elimination-reconstruction procedures. Bar is 50 μm. * p < 0.05 vs. WT or vs. Clo + WT
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Effect of FGF2 deficiency on BMDM apoptosis and polarization. a – c FGF2 deletion increased BMDM apoptosis. a Apoptosis in BMDM deprived of FBS for 24 h was assessed by flow cytometry ( n = 4). b - c Percentage of PI + Annexin V + and PI- Annexin V + BMDM after starvation. d - k FGF2 deletion in BMDM promoted M1 polarization. d - g Flow cytometric analysis of macrophage markers in BMDM treated with LPS or IL4, including CD86, iNOS, CD206, and Arg1 ( n = 3). h - k The levels of CD86, iNOS, CD206 and Arg1 in BMDM after treatment with LPS or IL4. N represents no treatment; * p < 0.05, vs. WT; Ψ p < 0.05, vs. N + WT; Ω p < 0.05, vs. N + FGF2 KO
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Deficiency of FGF2 in BMDM resulted in the upregulation of M1 markers and proinflammatory cytokine expression and increased nuclear translocation of NF-KB p65. a - j BMDM were treated with LPS or IL4, and real-time PCR was used to determine the expression of M1 and M2 markers and cytokines ( n = 3). k , l P65 nuclear translocation was detected and quantitatively analyzed by immunofluorescence ( n = 3). m The expression of MMP9 in WT and FGF2 KO BMDM treated with LPS or IL4 were determined with real-time PCR. Bar in the first three rows is 50 μm, while bar in the fourth rows is 20 μm, * p < 0.05 vs. WT; Ψ p < 0.05 vs. N + WT; Ω p < 0.05 vs. N + KO
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Transcriptome sequencing was used to analyze BMDM from WT and FGF2 KO mice treated with or without LPS. a Heat map of DEGs in BMDM from WT and FGF2 KO mice stimulated with or without LPS (Average TPM each group, n = 3). b KEGG enrichment analysis identified the top 20 altered signaling pathways in the four groups (WT, FGF2 KO, WT + LPS, and FGF2 KO + LPS). c Construction of a regulatory network to modulate PPI involving FGF2 and LPS using DEGs. d KEGG pathway network based on similarity in gene expression profiles. e Volcano plot showing DEGs of WT + LPS and FGF2 KO + LPS. f Heat map of DEGs in BMDM of WT + LPS and FGF2 KO + LPS. g Top 20 KEGG pathways for DEGs in BMDM from WT + LPS and FGF2 KO + LPS. h KEGG pathway annotation of differentially expressed genes between WT + LPS and FGF2 KO + LPS. i , j Differently regulated pathways in the GSEA. k Inflammation and cytokine gene heat map for WT + LPS and FGF2 KO + LPS. l , m TPM changes in the gene groups compared to WT weights. * p < 0.05 vs. WT; Δ p < 0.05 vs. WT + LPS; # p < 0.05 vs. FGF2 KO
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FGF2 deletion in macrophages aggravates lung injury in CLP mice. a - c Clodronate liposomes were injected intravenously to deplete macrophages, followed by quantification of F4/80 + spleen cells using flow cytometry to measure macrophage clearance. * p < 0.05 vs. control. d - f Levels of the inflammatory cytokines IL1β, TNFα, and IL6 in bronchoalveolar lavage fluid (BALF) from the four groups were quantified via ELISA. The four groups were Clo + WT + Sham, Clo + WT + CLP, Clo + FGF2 KO + Sham, and Clo + FGF2 KO + CLP. g The lung wet-to-dry weight ratios were measured in the four groups. h - i The total cells and total protein in BALF were evaluated in the four groups. j The OD value of Evans blue in the lung tissues from the four groups. k - l HE staining of lung tissue from the four groups and their Smith score. (m-o) Blood gas analysis was performed using abdominal aortic blood from the four groups. Bar is 250 μm. * p < 0.05 vs. WT; Δ p < 0.05 vs. WT + LPS; # p < 0.05 vs. FGF2 KO
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Mice reconstituted with FGF2 KO macrophages and subjected to CLP demonstrate increased M1 polarization in lung tissue. a - f The presence and levels of CD206, CD86, and F4/80 markers on macrophages within lung tissue were identified and quantitatively assessed using immunofluorescence staining. Bar is 20 μm. * p < 0.05, vs. WT; Δ p < 0.05 vs. WT + LPS; # p < 0.05 vs. FGF2 KO
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Mice reconstituted with FGF2 KO macrophages and subjected to CLP exhibit increased proinflammatory gene expression and apoptosis. a - c The expression of the inflammatory factors CXCL1, IL1β, and IL6 in lung tissue was detected by real-time PCR ( n = 3 per group). d TUNEL staining was used to evaluate apoptosis in the lung tissue. e - f The apoptosis-associated genes BCL2 and Bax were identified by western blot analysis. Bar is 20 μm. * p < 0.05, vs. WT; Δ p < 0.05, vs. WT + LPS; # p < 0.05 vs. FGF2 KO
Index in PubMed under a CC BY license. PMID: 39436561