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leptin receptor  (Bioss)


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    Bioss leptin receptor
    a Representative images of calcein and xylenol orange double labeling of bone and quantification of MAR and BFR for MK deleted mice and their littermate controls ( n = 6 mice per group). Scale bar, 100 µm. b Representative immunostaining images of <t>LepR</t> (red) in the BM of MK deleted mice and their littermate controls. The quantification of LepR + cells is shown in the right ( n = 6 mice per group). Scale bar, 100 µm. <t>c</t> <t>Osteocalcin</t> concentration in the BM of MK deleted mice and their littermate controls, determined by ELISA ( n = 6 mice per group). d Osteocalcin concentration in the serum of MK deleted mice and their littermate controls, determined by ELISA ( n = 6 mice per group). e PINP in the serum of MK deleted mice and their littermate controls, determined by ELISA ( n = 6 mice per group). f , g Representative immunostaining images of OCN (red) in the EB ( f ) and TB ( g ) of MK deleted mice and their littermate controls. The quantification of OCN + cells is shown on the right graphs ( n = 6 mice per group). Scale bar, 100 µm. h Quantitative biomechanical analysis of femora (peak load and stiffness) from MK deleted mice and their littermate controls ( n = 6 mice per group). i HE staining demonstrating metaphyseal bone and BM sections for MKs (black arrowheads at 48 h post-irradiation ( n = 6 mice per group). Scale bar, 100 µm. j Fluorescent images of mouse femoral bone. Lepr + cells (red) 7 days and 1 month after irradiation ( n = 6 mice per group). k Representative flow cytometry plots and quantification of percent MKs (CD41 + CD42d + ) and LepR + SSCs (Lepr + CD45 − CD31 − Ter119 − ) 7 days after irradiation ( n = 6 mice per group). Data on graphs are shown as mean ± SD. An unpaired two-tailed t -test was used to analyze the data in a – h and one-way ANOVA was used to analyze the data in j and k . * P < 0.05, ** P < 0.01 and *** P < 0.001. For all panels in this figure, data are representative of three independent experiments.
    Leptin Receptor, supplied by Bioss, used in various techniques. Bioz Stars score: 93/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 93 stars, based on 4 article reviews
    leptin receptor - by Bioz Stars, 2026-03
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    Images

    1) Product Images from "Megakaryocytic TGFβ1 orchestrates osteogenesis of LepR + SSCs to alleviate radiation-induced bone loss"

    Article Title: Megakaryocytic TGFβ1 orchestrates osteogenesis of LepR + SSCs to alleviate radiation-induced bone loss

    Journal: Experimental & Molecular Medicine

    doi: 10.1038/s12276-025-01612-z

    a Representative images of calcein and xylenol orange double labeling of bone and quantification of MAR and BFR for MK deleted mice and their littermate controls ( n = 6 mice per group). Scale bar, 100 µm. b Representative immunostaining images of LepR (red) in the BM of MK deleted mice and their littermate controls. The quantification of LepR + cells is shown in the right ( n = 6 mice per group). Scale bar, 100 µm. c Osteocalcin concentration in the BM of MK deleted mice and their littermate controls, determined by ELISA ( n = 6 mice per group). d Osteocalcin concentration in the serum of MK deleted mice and their littermate controls, determined by ELISA ( n = 6 mice per group). e PINP in the serum of MK deleted mice and their littermate controls, determined by ELISA ( n = 6 mice per group). f , g Representative immunostaining images of OCN (red) in the EB ( f ) and TB ( g ) of MK deleted mice and their littermate controls. The quantification of OCN + cells is shown on the right graphs ( n = 6 mice per group). Scale bar, 100 µm. h Quantitative biomechanical analysis of femora (peak load and stiffness) from MK deleted mice and their littermate controls ( n = 6 mice per group). i HE staining demonstrating metaphyseal bone and BM sections for MKs (black arrowheads at 48 h post-irradiation ( n = 6 mice per group). Scale bar, 100 µm. j Fluorescent images of mouse femoral bone. Lepr + cells (red) 7 days and 1 month after irradiation ( n = 6 mice per group). k Representative flow cytometry plots and quantification of percent MKs (CD41 + CD42d + ) and LepR + SSCs (Lepr + CD45 − CD31 − Ter119 − ) 7 days after irradiation ( n = 6 mice per group). Data on graphs are shown as mean ± SD. An unpaired two-tailed t -test was used to analyze the data in a – h and one-way ANOVA was used to analyze the data in j and k . * P < 0.05, ** P < 0.01 and *** P < 0.001. For all panels in this figure, data are representative of three independent experiments.
    Figure Legend Snippet: a Representative images of calcein and xylenol orange double labeling of bone and quantification of MAR and BFR for MK deleted mice and their littermate controls ( n = 6 mice per group). Scale bar, 100 µm. b Representative immunostaining images of LepR (red) in the BM of MK deleted mice and their littermate controls. The quantification of LepR + cells is shown in the right ( n = 6 mice per group). Scale bar, 100 µm. c Osteocalcin concentration in the BM of MK deleted mice and their littermate controls, determined by ELISA ( n = 6 mice per group). d Osteocalcin concentration in the serum of MK deleted mice and their littermate controls, determined by ELISA ( n = 6 mice per group). e PINP in the serum of MK deleted mice and their littermate controls, determined by ELISA ( n = 6 mice per group). f , g Representative immunostaining images of OCN (red) in the EB ( f ) and TB ( g ) of MK deleted mice and their littermate controls. The quantification of OCN + cells is shown on the right graphs ( n = 6 mice per group). Scale bar, 100 µm. h Quantitative biomechanical analysis of femora (peak load and stiffness) from MK deleted mice and their littermate controls ( n = 6 mice per group). i HE staining demonstrating metaphyseal bone and BM sections for MKs (black arrowheads at 48 h post-irradiation ( n = 6 mice per group). Scale bar, 100 µm. j Fluorescent images of mouse femoral bone. Lepr + cells (red) 7 days and 1 month after irradiation ( n = 6 mice per group). k Representative flow cytometry plots and quantification of percent MKs (CD41 + CD42d + ) and LepR + SSCs (Lepr + CD45 − CD31 − Ter119 − ) 7 days after irradiation ( n = 6 mice per group). Data on graphs are shown as mean ± SD. An unpaired two-tailed t -test was used to analyze the data in a – h and one-way ANOVA was used to analyze the data in j and k . * P < 0.05, ** P < 0.01 and *** P < 0.001. For all panels in this figure, data are representative of three independent experiments.

    Techniques Used: Labeling, Immunostaining, Concentration Assay, Enzyme-linked Immunosorbent Assay, Staining, Irradiation, Flow Cytometry, Two Tailed Test

    a Left: representative micro-CT images of longitudinal section femurs, cross-sectional view of the distal femurs and reconstructed trabecular structure of the region of interest from TGFβ1 MKΔ/Δ mice and their littermate controls (TGFβ1 fl/fl mice). Right: quantitative micro-CT analysis of the TB fraction (BV/TV, Tb.N, Tb.Th, Tb.Sp and Ct.Th) in TGFβ1 MKΔ/Δ mice and their littermate controls (TGFβ1 fl/fl mice) ( n = 6 mice per group). b LepR + SSCs were induced in osteogenic differentiation medium with or without MKs or (pretreated TGFβ type I receptor inhibitor SB431542) from wild-type (WT) mice after 14 days. Representative alkaline phosphatase staining images (left) and quantification of the activity of alkaline phosphatase was calculated (right) ( n = 6 per group). c LepR + SSCs were induced in osteogenic differentiation medium with or without MKs or (pretreated TGFβ type I receptor inhibitor SB431542) from WT mice after 21 days. Representative alizarin red staining images (left) and quantification of matrix mineralization was calculated (right) ( n = 6 per group). d LepR + SSCs were induced in adipogenic differentiation medium with or without MKs or (pretreated TGFβ type I receptor inhibitor SB431542) from WT mice after 21 days. Representative Oil O staining images (left) and the quantification of area was calculated (right) ( n = 6 per group). e qPCR analysis of the expression of Osterix , Runx2 , Adipoq and PPARγ in LepR + SSCs with or without MKs or (pretreated TGFβ type I receptor inhibitor SB431542) from WT mice after 7 days ( n = 3 per group). f LepR + SSCs were induced in osteogenic differentiation medium with or without MKs from the BM of TGFβ1 MKΔ/Δ and TGFβ1 fl/fl mice after 14 days. Representative alkaline phosphatase staining images and quantification of the activity of alkaline phosphatase was calculated ( n = 6 per group). g LepR + SSCs were induced in osteogenic differentiation medium with or without MKs from the BM of TGFβ1 MKΔ/Δ and TGFβ1 fl/fl mice after 21 days. Representative alizarin red staining images and quantification of matrix mineralization was calculated ( n = 6 per group). Data on graphs are shown as mean ± SD. One-way ANOVA was used to analyze the data in a – e and an unpaired two-tailed t -test was used to analyze the data in f and g . * P < 0.05, ** P < 0.01 and *** P < 0.001. For all panels in this figure, data are representative of three independent experiments.
    Figure Legend Snippet: a Left: representative micro-CT images of longitudinal section femurs, cross-sectional view of the distal femurs and reconstructed trabecular structure of the region of interest from TGFβ1 MKΔ/Δ mice and their littermate controls (TGFβ1 fl/fl mice). Right: quantitative micro-CT analysis of the TB fraction (BV/TV, Tb.N, Tb.Th, Tb.Sp and Ct.Th) in TGFβ1 MKΔ/Δ mice and their littermate controls (TGFβ1 fl/fl mice) ( n = 6 mice per group). b LepR + SSCs were induced in osteogenic differentiation medium with or without MKs or (pretreated TGFβ type I receptor inhibitor SB431542) from wild-type (WT) mice after 14 days. Representative alkaline phosphatase staining images (left) and quantification of the activity of alkaline phosphatase was calculated (right) ( n = 6 per group). c LepR + SSCs were induced in osteogenic differentiation medium with or without MKs or (pretreated TGFβ type I receptor inhibitor SB431542) from WT mice after 21 days. Representative alizarin red staining images (left) and quantification of matrix mineralization was calculated (right) ( n = 6 per group). d LepR + SSCs were induced in adipogenic differentiation medium with or without MKs or (pretreated TGFβ type I receptor inhibitor SB431542) from WT mice after 21 days. Representative Oil O staining images (left) and the quantification of area was calculated (right) ( n = 6 per group). e qPCR analysis of the expression of Osterix , Runx2 , Adipoq and PPARγ in LepR + SSCs with or without MKs or (pretreated TGFβ type I receptor inhibitor SB431542) from WT mice after 7 days ( n = 3 per group). f LepR + SSCs were induced in osteogenic differentiation medium with or without MKs from the BM of TGFβ1 MKΔ/Δ and TGFβ1 fl/fl mice after 14 days. Representative alkaline phosphatase staining images and quantification of the activity of alkaline phosphatase was calculated ( n = 6 per group). g LepR + SSCs were induced in osteogenic differentiation medium with or without MKs from the BM of TGFβ1 MKΔ/Δ and TGFβ1 fl/fl mice after 21 days. Representative alizarin red staining images and quantification of matrix mineralization was calculated ( n = 6 per group). Data on graphs are shown as mean ± SD. One-way ANOVA was used to analyze the data in a – e and an unpaired two-tailed t -test was used to analyze the data in f and g . * P < 0.05, ** P < 0.01 and *** P < 0.001. For all panels in this figure, data are representative of three independent experiments.

    Techniques Used: Micro-CT, Staining, Activity Assay, Expressing, Two Tailed Test

    a RNA-seq analysis revealed changes in gene expression in LepR + SSCs co-cultured with MKs ( n = 3 each). b qPCR analysis of the expressions of S mad2 and Slc39a14 in LepR + SSCs, with or without MKs, from WT mice ( n = 6 per group). c Western blotting analysis of the expression of Smad2 and Slc39a14 in LepR + SSCs, with or without MKs, from WT mice ( n = 3 per group). d Representative immunostaining images of Smad2 (red) and Slc39a14 (green) in LepR + SSCs, with or without MKs, from the BM of TGFβ1 MKΔ/Δ and TGFβ1 fl/fl mice ( n = 6 per group). Scale bar, 100 µm. e Colocalization of Smad2 (red) with Slc39a14 (green) in LepR + SSCs, with or without MKs, from the BM of TGFβ1 MKΔ/Δ and TGFβ1 fl/fl mice ( n = 6 per group). Ctrl, control. f A schematic representation of the neural network model of Smad2 binding to the promoter region of Slc39a14, predicted by AlphaFold 3. g A plot of the predicted aligned error of the complex predicted by AlphaFold 3 (pTM + ipTM = 0.91). h A plot of the binding site and amino acid residues of Smad2–Slc39a14 analyzed by PyMol. i Dual-luciferase assays of 293T cotransfected with WT or mutated Slc39a14 (LUC), combined with pcDNA3.1-Smad2 or pcDNA3.1 vetor. j ChIP assay of Smad2 binding to Slc39a14 promoters in LepR + SSCs transfected with pcDNA3.1-Smad2 or pcDNA3.1. Immunoprecipitated DNA and the input DNA were detected by PCR. Primer sequences were designed for Slc39a14 promoter regions located in the promoter region of the Slc39a14 gene, with IgG as a negative control. Data on graphs are shown as mean ± SD. An unpaired two-tailed t -test was used to analyze the data in b and i . * P < 0.05, ** P < 0.01 and *** P < 0.001. For all panels in this figure, data are representative of three independent experiments.
    Figure Legend Snippet: a RNA-seq analysis revealed changes in gene expression in LepR + SSCs co-cultured with MKs ( n = 3 each). b qPCR analysis of the expressions of S mad2 and Slc39a14 in LepR + SSCs, with or without MKs, from WT mice ( n = 6 per group). c Western blotting analysis of the expression of Smad2 and Slc39a14 in LepR + SSCs, with or without MKs, from WT mice ( n = 3 per group). d Representative immunostaining images of Smad2 (red) and Slc39a14 (green) in LepR + SSCs, with or without MKs, from the BM of TGFβ1 MKΔ/Δ and TGFβ1 fl/fl mice ( n = 6 per group). Scale bar, 100 µm. e Colocalization of Smad2 (red) with Slc39a14 (green) in LepR + SSCs, with or without MKs, from the BM of TGFβ1 MKΔ/Δ and TGFβ1 fl/fl mice ( n = 6 per group). Ctrl, control. f A schematic representation of the neural network model of Smad2 binding to the promoter region of Slc39a14, predicted by AlphaFold 3. g A plot of the predicted aligned error of the complex predicted by AlphaFold 3 (pTM + ipTM = 0.91). h A plot of the binding site and amino acid residues of Smad2–Slc39a14 analyzed by PyMol. i Dual-luciferase assays of 293T cotransfected with WT or mutated Slc39a14 (LUC), combined with pcDNA3.1-Smad2 or pcDNA3.1 vetor. j ChIP assay of Smad2 binding to Slc39a14 promoters in LepR + SSCs transfected with pcDNA3.1-Smad2 or pcDNA3.1. Immunoprecipitated DNA and the input DNA were detected by PCR. Primer sequences were designed for Slc39a14 promoter regions located in the promoter region of the Slc39a14 gene, with IgG as a negative control. Data on graphs are shown as mean ± SD. An unpaired two-tailed t -test was used to analyze the data in b and i . * P < 0.05, ** P < 0.01 and *** P < 0.001. For all panels in this figure, data are representative of three independent experiments.

    Techniques Used: RNA Sequencing, Gene Expression, Cell Culture, Western Blot, Expressing, Immunostaining, Control, Binding Assay, Luciferase, Transfection, Immunoprecipitation, Negative Control, Two Tailed Test

    a Serum zinc concentration in mice 4 weeks after irradiation with administration of TPO or vehicle ( n = 6 per group). b Representative fluozin-3 images and quantitative analysis of LepR + SSCs, with or without MKs, after irradiation ( n = 6 per group). c Representative immunostaining images of Slc39a14 (green) in LepR + SSCs, with or without MKs, after irradiation ( n = 6 per group). Scale bar, 100 µm. d KEGG enrichment analysis of upregulated pathways in LepR + SSCs after irradiation. e GO enrichment analysis of downregulated functions in LepR + SSCs after co-culture with MKs. The top ten enriched GO terms ( P < 0.05) are shown. f Western blotting analysis of the expression of Slc39a14, PTP1B, p-eIF2α, ATF4 and CHOP in LepR + SSCs after co-culture with MKs ( n = 3 per group). g Representative immunostaining images of CHOP (green) in LepR + SSCs, with or without MKs, after irradiation ( n = 6 per group). Scale bar, 100 µm. h Transmission electron microscopy images of LepR + SSCs after irradiation co-culture with MKs ( n = 3 per group). Data on graphs are shown as mean ± SD. One-way ANOVA was used to analyze the data in a – d and g . * P < 0.05, ** P < 0.01 and *** P < 0.001. For all panels in this figure, data are representative of three independent experiments.
    Figure Legend Snippet: a Serum zinc concentration in mice 4 weeks after irradiation with administration of TPO or vehicle ( n = 6 per group). b Representative fluozin-3 images and quantitative analysis of LepR + SSCs, with or without MKs, after irradiation ( n = 6 per group). c Representative immunostaining images of Slc39a14 (green) in LepR + SSCs, with or without MKs, after irradiation ( n = 6 per group). Scale bar, 100 µm. d KEGG enrichment analysis of upregulated pathways in LepR + SSCs after irradiation. e GO enrichment analysis of downregulated functions in LepR + SSCs after co-culture with MKs. The top ten enriched GO terms ( P < 0.05) are shown. f Western blotting analysis of the expression of Slc39a14, PTP1B, p-eIF2α, ATF4 and CHOP in LepR + SSCs after co-culture with MKs ( n = 3 per group). g Representative immunostaining images of CHOP (green) in LepR + SSCs, with or without MKs, after irradiation ( n = 6 per group). Scale bar, 100 µm. h Transmission electron microscopy images of LepR + SSCs after irradiation co-culture with MKs ( n = 3 per group). Data on graphs are shown as mean ± SD. One-way ANOVA was used to analyze the data in a – d and g . * P < 0.05, ** P < 0.01 and *** P < 0.001. For all panels in this figure, data are representative of three independent experiments.

    Techniques Used: Concentration Assay, Irradiation, Immunostaining, Co-Culture Assay, Western Blot, Expressing, Transmission Assay, Electron Microscopy

    a Representative immunostaining images of LepR (red) and PTP1B (green) in the BM of irradiated mice ( n = 6 mice per group). Scale bar, 100 µm. b Representative immunostaining images of LepR (red) and PTP1B (green) in the BM of MK deleted mice and their littermate controls after irradiation ( n = 6 mice per group). Scale bar, 100 µm. c Representative immunostaining images of PTP1B in LepR + cells, with or without, MKs from the BM of TGFβ1 MKΔ/Δ and TGFβ1 fl/fl mice after irradiation ( n = 6 per group). Scale bar, 100 µm. d Western blotting analysis of the expression of PTP1B and p-Stat3 in LepR + SSCs after co-culture with MKs ( n = 3 per group), inh = inhibitor. e Western blotting analysis of the expression of PTP1B and p-Stat3 in LepR + SSCs after co-culture with MKs from the BM of TGFβ1 MKΔ/Δ and TGFβ1 fl/fl mice ( n = 3 per group). Data on graphs are shown as mean ± SD. One-way ANOVA was used to analyze the data in c and d . * P < 0.05, ** P < 0.01 and *** P < 0.001. For all panels in this figure, data are representative of three independent experiments.
    Figure Legend Snippet: a Representative immunostaining images of LepR (red) and PTP1B (green) in the BM of irradiated mice ( n = 6 mice per group). Scale bar, 100 µm. b Representative immunostaining images of LepR (red) and PTP1B (green) in the BM of MK deleted mice and their littermate controls after irradiation ( n = 6 mice per group). Scale bar, 100 µm. c Representative immunostaining images of PTP1B in LepR + cells, with or without, MKs from the BM of TGFβ1 MKΔ/Δ and TGFβ1 fl/fl mice after irradiation ( n = 6 per group). Scale bar, 100 µm. d Western blotting analysis of the expression of PTP1B and p-Stat3 in LepR + SSCs after co-culture with MKs ( n = 3 per group), inh = inhibitor. e Western blotting analysis of the expression of PTP1B and p-Stat3 in LepR + SSCs after co-culture with MKs from the BM of TGFβ1 MKΔ/Δ and TGFβ1 fl/fl mice ( n = 3 per group). Data on graphs are shown as mean ± SD. One-way ANOVA was used to analyze the data in c and d . * P < 0.05, ** P < 0.01 and *** P < 0.001. For all panels in this figure, data are representative of three independent experiments.

    Techniques Used: Immunostaining, Irradiation, Western Blot, Expressing, Co-Culture Assay

    a Representative micro-CT images of longitudinal section femurs, cross-sectional view of the distal femurs and reconstructed trabecular structure of the region of interest from mice injected with TPO or vehicle after irradiation ( n = 6 mice per group). b Quantitative micro-CT analysis of the TB fraction (BV/TV, Tb.N, Tb.Th, Tb.Sp, BMD and Ct.Th) in mice injected with TPO or vehicle after irradiation ( n = 6 mice per group). c Representative immunostaining images of OCN (red) in the TB and EB of the mice injected with TPO or vehicle after irradiation. The quantification of OCN cells is shown on the right ( n = 6 mice per group). Scale bar, 100 µm. d HE staining demonstrating B.Ar/T.Ar (bone area/total area) and the presence of MKs in the osteogenic niche of control or irradiated mice 8 weeks after injection with TPO ( n = 6 mice per group). Scale bar, 100 µm. e Von Kossa staining showing mineralization of bone matrix in control or irradiated mice 8 weeks after injected with TPO ( n = 6 mice per group). Scale bar, 1 mm. f Representative immunostaining images of perilipin (red) and osteopontin (OPN, green) in the BM of irradiation mice ( n = 6 mice per group). Scale bar, 100 µm. g Colocalization of LepR (red) with Slc39a14 (green) in the BM of irradiation mice ( n = 6 mice per group). Scale bar, 100 µm. Data on graphs are shown as mean ± SD. One-way ANOVA was used to analyze the data in b – g . * P < 0.05, ** P < 0.01 and *** P < 0.001. For all panels in this figure, data are representative of three independent experiments.
    Figure Legend Snippet: a Representative micro-CT images of longitudinal section femurs, cross-sectional view of the distal femurs and reconstructed trabecular structure of the region of interest from mice injected with TPO or vehicle after irradiation ( n = 6 mice per group). b Quantitative micro-CT analysis of the TB fraction (BV/TV, Tb.N, Tb.Th, Tb.Sp, BMD and Ct.Th) in mice injected with TPO or vehicle after irradiation ( n = 6 mice per group). c Representative immunostaining images of OCN (red) in the TB and EB of the mice injected with TPO or vehicle after irradiation. The quantification of OCN cells is shown on the right ( n = 6 mice per group). Scale bar, 100 µm. d HE staining demonstrating B.Ar/T.Ar (bone area/total area) and the presence of MKs in the osteogenic niche of control or irradiated mice 8 weeks after injection with TPO ( n = 6 mice per group). Scale bar, 100 µm. e Von Kossa staining showing mineralization of bone matrix in control or irradiated mice 8 weeks after injected with TPO ( n = 6 mice per group). Scale bar, 1 mm. f Representative immunostaining images of perilipin (red) and osteopontin (OPN, green) in the BM of irradiation mice ( n = 6 mice per group). Scale bar, 100 µm. g Colocalization of LepR (red) with Slc39a14 (green) in the BM of irradiation mice ( n = 6 mice per group). Scale bar, 100 µm. Data on graphs are shown as mean ± SD. One-way ANOVA was used to analyze the data in b – g . * P < 0.05, ** P < 0.01 and *** P < 0.001. For all panels in this figure, data are representative of three independent experiments.

    Techniques Used: Micro-CT, Injection, Irradiation, Immunostaining, Staining, Control

    a Representative micro-CT images of longitudinal section femurs, cross-sectional view of the distal femurs and reconstructed trabecular structure of the region of interest from Slc39a14 leprΔ/Δ mice and their littermate controls (Slc39a14 fl/fl mice) ( n = 6 mice per group). b Quantitative micro-CT analysis of the TB fraction (BV/TV, Tb.N, Tb.Th, Tb.Sp, BMD and Ct.Th) in Slc39a14 leprΔ/Δ mice and their littermate controls (Slc39a14 fl/fl mice) ( n = 6 mice per group). c Quantitative biomechanical analysis of femora (peak load and stiffness) from MK deleted mice and their littermate controls (Slc39a14 fl/fl mice) ( n = 6 mice per group). d Serum zinc concentration in Slc39a14 leprΔ/Δ mice and their littermate controls (Slc39a14 fl/fl mice) ( n = 6 per group). e Von Kossa staining showing mineralization of bone matrix in Slc39a14 leprΔ/Δ mice and their littermate controls (Slc39a14 fl/fl mice) ( n = 6 per group). f Representative micro-CT images of longitudinal section femurs, cross-sectional view of the distal femurs and reconstructed trabecular structure of the region of interest from Slc39a14 leprΔ/Δ mice injected with TPO or vehicle after irradiation ( n = 6 mice per group). g Representative immunostaining images of OCN (red) in the TB and EB of the Slc39a14 leprΔ/Δ mice injected with TPO or vehicle after irradiation. The quantification of OCN cells is shown on the right ( n = 6 mice per group). Scale bar, 100 µm. h Representative immunostaining images of LepR (red) and PTP1B (green) in the BM of the Slc39a14 leprΔ/Δ mice injected with TPO or vehicle after irradiation. Scale bar, 100 µm. i Colocalization of LepR (red) with Ki67 (green) in the BM of the Slc39a14 leprΔ/Δ mice injected with TPO or vehicle after irradiation ( n = 6 mice per group). Scale bar, 100 µm. j Representative immunostaining images of TUNEL (green) in the BM of the Slc39a14 leprΔ/Δ mice injected with TPO or vehicle after irradiation. The quantification of tunel positive cells is shown on the right ( n = 6 mice per group). Scale bar, 100 µm. Data on graphs are shown as mean ± SD. An unpaired two-tailed t -test was used to analyze the data in b – d , g , i and j . * P < 0.05, ** P < 0.01 and *** P < 0.001. For all panels in this figure, data are representative of three independent experiments.
    Figure Legend Snippet: a Representative micro-CT images of longitudinal section femurs, cross-sectional view of the distal femurs and reconstructed trabecular structure of the region of interest from Slc39a14 leprΔ/Δ mice and their littermate controls (Slc39a14 fl/fl mice) ( n = 6 mice per group). b Quantitative micro-CT analysis of the TB fraction (BV/TV, Tb.N, Tb.Th, Tb.Sp, BMD and Ct.Th) in Slc39a14 leprΔ/Δ mice and their littermate controls (Slc39a14 fl/fl mice) ( n = 6 mice per group). c Quantitative biomechanical analysis of femora (peak load and stiffness) from MK deleted mice and their littermate controls (Slc39a14 fl/fl mice) ( n = 6 mice per group). d Serum zinc concentration in Slc39a14 leprΔ/Δ mice and their littermate controls (Slc39a14 fl/fl mice) ( n = 6 per group). e Von Kossa staining showing mineralization of bone matrix in Slc39a14 leprΔ/Δ mice and their littermate controls (Slc39a14 fl/fl mice) ( n = 6 per group). f Representative micro-CT images of longitudinal section femurs, cross-sectional view of the distal femurs and reconstructed trabecular structure of the region of interest from Slc39a14 leprΔ/Δ mice injected with TPO or vehicle after irradiation ( n = 6 mice per group). g Representative immunostaining images of OCN (red) in the TB and EB of the Slc39a14 leprΔ/Δ mice injected with TPO or vehicle after irradiation. The quantification of OCN cells is shown on the right ( n = 6 mice per group). Scale bar, 100 µm. h Representative immunostaining images of LepR (red) and PTP1B (green) in the BM of the Slc39a14 leprΔ/Δ mice injected with TPO or vehicle after irradiation. Scale bar, 100 µm. i Colocalization of LepR (red) with Ki67 (green) in the BM of the Slc39a14 leprΔ/Δ mice injected with TPO or vehicle after irradiation ( n = 6 mice per group). Scale bar, 100 µm. j Representative immunostaining images of TUNEL (green) in the BM of the Slc39a14 leprΔ/Δ mice injected with TPO or vehicle after irradiation. The quantification of tunel positive cells is shown on the right ( n = 6 mice per group). Scale bar, 100 µm. Data on graphs are shown as mean ± SD. An unpaired two-tailed t -test was used to analyze the data in b – d , g , i and j . * P < 0.05, ** P < 0.01 and *** P < 0.001. For all panels in this figure, data are representative of three independent experiments.

    Techniques Used: Micro-CT, Concentration Assay, Staining, Injection, Irradiation, Immunostaining, TUNEL Assay, Two Tailed Test



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    93
    Bioss leptin receptor
    a Representative images of calcein and xylenol orange double labeling of bone and quantification of MAR and BFR for MK deleted mice and their littermate controls ( n = 6 mice per group). Scale bar, 100 µm. b Representative immunostaining images of <t>LepR</t> (red) in the BM of MK deleted mice and their littermate controls. The quantification of LepR + cells is shown in the right ( n = 6 mice per group). Scale bar, 100 µm. <t>c</t> <t>Osteocalcin</t> concentration in the BM of MK deleted mice and their littermate controls, determined by ELISA ( n = 6 mice per group). d Osteocalcin concentration in the serum of MK deleted mice and their littermate controls, determined by ELISA ( n = 6 mice per group). e PINP in the serum of MK deleted mice and their littermate controls, determined by ELISA ( n = 6 mice per group). f , g Representative immunostaining images of OCN (red) in the EB ( f ) and TB ( g ) of MK deleted mice and their littermate controls. The quantification of OCN + cells is shown on the right graphs ( n = 6 mice per group). Scale bar, 100 µm. h Quantitative biomechanical analysis of femora (peak load and stiffness) from MK deleted mice and their littermate controls ( n = 6 mice per group). i HE staining demonstrating metaphyseal bone and BM sections for MKs (black arrowheads at 48 h post-irradiation ( n = 6 mice per group). Scale bar, 100 µm. j Fluorescent images of mouse femoral bone. Lepr + cells (red) 7 days and 1 month after irradiation ( n = 6 mice per group). k Representative flow cytometry plots and quantification of percent MKs (CD41 + CD42d + ) and LepR + SSCs (Lepr + CD45 − CD31 − Ter119 − ) 7 days after irradiation ( n = 6 mice per group). Data on graphs are shown as mean ± SD. An unpaired two-tailed t -test was used to analyze the data in a – h and one-way ANOVA was used to analyze the data in j and k . * P < 0.05, ** P < 0.01 and *** P < 0.001. For all panels in this figure, data are representative of three independent experiments.
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    Bioss anti leptin receptor primary antibody
    a Representative images of calcein and xylenol orange double labeling of bone and quantification of MAR and BFR for MK deleted mice and their littermate controls ( n = 6 mice per group). Scale bar, 100 µm. b Representative immunostaining images of <t>LepR</t> (red) in the BM of MK deleted mice and their littermate controls. The quantification of LepR + cells is shown in the right ( n = 6 mice per group). Scale bar, 100 µm. <t>c</t> <t>Osteocalcin</t> concentration in the BM of MK deleted mice and their littermate controls, determined by ELISA ( n = 6 mice per group). d Osteocalcin concentration in the serum of MK deleted mice and their littermate controls, determined by ELISA ( n = 6 mice per group). e PINP in the serum of MK deleted mice and their littermate controls, determined by ELISA ( n = 6 mice per group). f , g Representative immunostaining images of OCN (red) in the EB ( f ) and TB ( g ) of MK deleted mice and their littermate controls. The quantification of OCN + cells is shown on the right graphs ( n = 6 mice per group). Scale bar, 100 µm. h Quantitative biomechanical analysis of femora (peak load and stiffness) from MK deleted mice and their littermate controls ( n = 6 mice per group). i HE staining demonstrating metaphyseal bone and BM sections for MKs (black arrowheads at 48 h post-irradiation ( n = 6 mice per group). Scale bar, 100 µm. j Fluorescent images of mouse femoral bone. Lepr + cells (red) 7 days and 1 month after irradiation ( n = 6 mice per group). k Representative flow cytometry plots and quantification of percent MKs (CD41 + CD42d + ) and LepR + SSCs (Lepr + CD45 − CD31 − Ter119 − ) 7 days after irradiation ( n = 6 mice per group). Data on graphs are shown as mean ± SD. An unpaired two-tailed t -test was used to analyze the data in a – h and one-way ANOVA was used to analyze the data in j and k . * P < 0.05, ** P < 0.01 and *** P < 0.001. For all panels in this figure, data are representative of three independent experiments.
    Anti Leptin Receptor Primary Antibody, supplied by Bioss, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    lepr  (Bioss)
    92
    Bioss lepr
    Age was not associated with any of the brain protein loads in the DS and DSAD groups. In the controls, we found associations between age and <t>LepR</t> ( p = 0.01), pSTAT3‐727 ( p = 0.04), and pSTAT3‐705 ( p = 0.002). DS, Down syndrome; DSAD, Down syndrome and Alzheimer's disease; <t>LepR,</t> <t>leptin</t> receptor; pSTAT3, phosphorylated STAT3.
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    Thermo Fisher leptin receptor polyclonal antibody pa1-053
    Age was not associated with any of the brain protein loads in the DS and DSAD groups. In the controls, we found associations between age and <t>LepR</t> ( p = 0.01), pSTAT3‐727 ( p = 0.04), and pSTAT3‐705 ( p = 0.002). DS, Down syndrome; DSAD, Down syndrome and Alzheimer's disease; <t>LepR,</t> <t>leptin</t> receptor; pSTAT3, phosphorylated STAT3.
    Leptin Receptor Polyclonal Antibody Pa1 053, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Boster Bio rabbit anti leptin receptor polyclonal antibody
    Age was not associated with any of the brain protein loads in the DS and DSAD groups. In the controls, we found associations between age and <t>LepR</t> ( p = 0.01), pSTAT3‐727 ( p = 0.04), and pSTAT3‐705 ( p = 0.002). DS, Down syndrome; DSAD, Down syndrome and Alzheimer's disease; <t>LepR,</t> <t>leptin</t> receptor; pSTAT3, phosphorylated STAT3.
    Rabbit Anti Leptin Receptor Polyclonal Antibody, supplied by Boster Bio, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Bioss anti lepr
    Age was not associated with any of the brain protein loads in the DS and DSAD groups. In the controls, we found associations between age and <t>LepR</t> ( p = 0.01), pSTAT3‐727 ( p = 0.04), and pSTAT3‐705 ( p = 0.002). DS, Down syndrome; DSAD, Down syndrome and Alzheimer's disease; <t>LepR,</t> <t>leptin</t> receptor; pSTAT3, phosphorylated STAT3.
    Anti Lepr, supplied by Bioss, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Bioss bs 0961r
    List of antibodies used in the study (IF—immunofluorescence staining; WB—Western blot analysis).
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    Bioss leptin receptor rabbit polyclonal antibody
    Viability of BME-UV1 bovine mammary epithelial cells treated for 24 h with different concentrations of chemerin, <t>leptin,</t> or adiponectin. Graphs present cell viability measured using the MTT assay. The viability of untreated control cells (0) was designated as 100%. Results are presented as means ± standard deviation of three independent experiments.
    Leptin Receptor Rabbit Polyclonal Antibody, supplied by Bioss, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    a Representative images of calcein and xylenol orange double labeling of bone and quantification of MAR and BFR for MK deleted mice and their littermate controls ( n = 6 mice per group). Scale bar, 100 µm. b Representative immunostaining images of LepR (red) in the BM of MK deleted mice and their littermate controls. The quantification of LepR + cells is shown in the right ( n = 6 mice per group). Scale bar, 100 µm. c Osteocalcin concentration in the BM of MK deleted mice and their littermate controls, determined by ELISA ( n = 6 mice per group). d Osteocalcin concentration in the serum of MK deleted mice and their littermate controls, determined by ELISA ( n = 6 mice per group). e PINP in the serum of MK deleted mice and their littermate controls, determined by ELISA ( n = 6 mice per group). f , g Representative immunostaining images of OCN (red) in the EB ( f ) and TB ( g ) of MK deleted mice and their littermate controls. The quantification of OCN + cells is shown on the right graphs ( n = 6 mice per group). Scale bar, 100 µm. h Quantitative biomechanical analysis of femora (peak load and stiffness) from MK deleted mice and their littermate controls ( n = 6 mice per group). i HE staining demonstrating metaphyseal bone and BM sections for MKs (black arrowheads at 48 h post-irradiation ( n = 6 mice per group). Scale bar, 100 µm. j Fluorescent images of mouse femoral bone. Lepr + cells (red) 7 days and 1 month after irradiation ( n = 6 mice per group). k Representative flow cytometry plots and quantification of percent MKs (CD41 + CD42d + ) and LepR + SSCs (Lepr + CD45 − CD31 − Ter119 − ) 7 days after irradiation ( n = 6 mice per group). Data on graphs are shown as mean ± SD. An unpaired two-tailed t -test was used to analyze the data in a – h and one-way ANOVA was used to analyze the data in j and k . * P < 0.05, ** P < 0.01 and *** P < 0.001. For all panels in this figure, data are representative of three independent experiments.

    Journal: Experimental & Molecular Medicine

    Article Title: Megakaryocytic TGFβ1 orchestrates osteogenesis of LepR + SSCs to alleviate radiation-induced bone loss

    doi: 10.1038/s12276-025-01612-z

    Figure Lengend Snippet: a Representative images of calcein and xylenol orange double labeling of bone and quantification of MAR and BFR for MK deleted mice and their littermate controls ( n = 6 mice per group). Scale bar, 100 µm. b Representative immunostaining images of LepR (red) in the BM of MK deleted mice and their littermate controls. The quantification of LepR + cells is shown in the right ( n = 6 mice per group). Scale bar, 100 µm. c Osteocalcin concentration in the BM of MK deleted mice and their littermate controls, determined by ELISA ( n = 6 mice per group). d Osteocalcin concentration in the serum of MK deleted mice and their littermate controls, determined by ELISA ( n = 6 mice per group). e PINP in the serum of MK deleted mice and their littermate controls, determined by ELISA ( n = 6 mice per group). f , g Representative immunostaining images of OCN (red) in the EB ( f ) and TB ( g ) of MK deleted mice and their littermate controls. The quantification of OCN + cells is shown on the right graphs ( n = 6 mice per group). Scale bar, 100 µm. h Quantitative biomechanical analysis of femora (peak load and stiffness) from MK deleted mice and their littermate controls ( n = 6 mice per group). i HE staining demonstrating metaphyseal bone and BM sections for MKs (black arrowheads at 48 h post-irradiation ( n = 6 mice per group). Scale bar, 100 µm. j Fluorescent images of mouse femoral bone. Lepr + cells (red) 7 days and 1 month after irradiation ( n = 6 mice per group). k Representative flow cytometry plots and quantification of percent MKs (CD41 + CD42d + ) and LepR + SSCs (Lepr + CD45 − CD31 − Ter119 − ) 7 days after irradiation ( n = 6 mice per group). Data on graphs are shown as mean ± SD. An unpaired two-tailed t -test was used to analyze the data in a – h and one-way ANOVA was used to analyze the data in j and k . * P < 0.05, ** P < 0.01 and *** P < 0.001. For all panels in this figure, data are representative of three independent experiments.

    Article Snippet: Briefly, the bone sections were incubated with primary antibodies against mouse osteocalcin (A20800, ABclonal), Ki67 (AF7617, R&D), PTP1B (bs-55182R, Bioss), Slc39a14 (A10413, ABclonal), leptin receptor (bs-0410R, Bioss), CHOP ( A21902 , ABclonal), F4/80 (30325, CST), TGFβ1 (ab313729, abcam), vWF (bsm-52775R, Bioss), osterix (ab209484, abcam) and Smad2 (A7699, ABclonal) overnight at 4 °C and incubated with secondary antibodies for 1 h at 37 °C.

    Techniques: Labeling, Immunostaining, Concentration Assay, Enzyme-linked Immunosorbent Assay, Staining, Irradiation, Flow Cytometry, Two Tailed Test

    a Left: representative micro-CT images of longitudinal section femurs, cross-sectional view of the distal femurs and reconstructed trabecular structure of the region of interest from TGFβ1 MKΔ/Δ mice and their littermate controls (TGFβ1 fl/fl mice). Right: quantitative micro-CT analysis of the TB fraction (BV/TV, Tb.N, Tb.Th, Tb.Sp and Ct.Th) in TGFβ1 MKΔ/Δ mice and their littermate controls (TGFβ1 fl/fl mice) ( n = 6 mice per group). b LepR + SSCs were induced in osteogenic differentiation medium with or without MKs or (pretreated TGFβ type I receptor inhibitor SB431542) from wild-type (WT) mice after 14 days. Representative alkaline phosphatase staining images (left) and quantification of the activity of alkaline phosphatase was calculated (right) ( n = 6 per group). c LepR + SSCs were induced in osteogenic differentiation medium with or without MKs or (pretreated TGFβ type I receptor inhibitor SB431542) from WT mice after 21 days. Representative alizarin red staining images (left) and quantification of matrix mineralization was calculated (right) ( n = 6 per group). d LepR + SSCs were induced in adipogenic differentiation medium with or without MKs or (pretreated TGFβ type I receptor inhibitor SB431542) from WT mice after 21 days. Representative Oil O staining images (left) and the quantification of area was calculated (right) ( n = 6 per group). e qPCR analysis of the expression of Osterix , Runx2 , Adipoq and PPARγ in LepR + SSCs with or without MKs or (pretreated TGFβ type I receptor inhibitor SB431542) from WT mice after 7 days ( n = 3 per group). f LepR + SSCs were induced in osteogenic differentiation medium with or without MKs from the BM of TGFβ1 MKΔ/Δ and TGFβ1 fl/fl mice after 14 days. Representative alkaline phosphatase staining images and quantification of the activity of alkaline phosphatase was calculated ( n = 6 per group). g LepR + SSCs were induced in osteogenic differentiation medium with or without MKs from the BM of TGFβ1 MKΔ/Δ and TGFβ1 fl/fl mice after 21 days. Representative alizarin red staining images and quantification of matrix mineralization was calculated ( n = 6 per group). Data on graphs are shown as mean ± SD. One-way ANOVA was used to analyze the data in a – e and an unpaired two-tailed t -test was used to analyze the data in f and g . * P < 0.05, ** P < 0.01 and *** P < 0.001. For all panels in this figure, data are representative of three independent experiments.

    Journal: Experimental & Molecular Medicine

    Article Title: Megakaryocytic TGFβ1 orchestrates osteogenesis of LepR + SSCs to alleviate radiation-induced bone loss

    doi: 10.1038/s12276-025-01612-z

    Figure Lengend Snippet: a Left: representative micro-CT images of longitudinal section femurs, cross-sectional view of the distal femurs and reconstructed trabecular structure of the region of interest from TGFβ1 MKΔ/Δ mice and their littermate controls (TGFβ1 fl/fl mice). Right: quantitative micro-CT analysis of the TB fraction (BV/TV, Tb.N, Tb.Th, Tb.Sp and Ct.Th) in TGFβ1 MKΔ/Δ mice and their littermate controls (TGFβ1 fl/fl mice) ( n = 6 mice per group). b LepR + SSCs were induced in osteogenic differentiation medium with or without MKs or (pretreated TGFβ type I receptor inhibitor SB431542) from wild-type (WT) mice after 14 days. Representative alkaline phosphatase staining images (left) and quantification of the activity of alkaline phosphatase was calculated (right) ( n = 6 per group). c LepR + SSCs were induced in osteogenic differentiation medium with or without MKs or (pretreated TGFβ type I receptor inhibitor SB431542) from WT mice after 21 days. Representative alizarin red staining images (left) and quantification of matrix mineralization was calculated (right) ( n = 6 per group). d LepR + SSCs were induced in adipogenic differentiation medium with or without MKs or (pretreated TGFβ type I receptor inhibitor SB431542) from WT mice after 21 days. Representative Oil O staining images (left) and the quantification of area was calculated (right) ( n = 6 per group). e qPCR analysis of the expression of Osterix , Runx2 , Adipoq and PPARγ in LepR + SSCs with or without MKs or (pretreated TGFβ type I receptor inhibitor SB431542) from WT mice after 7 days ( n = 3 per group). f LepR + SSCs were induced in osteogenic differentiation medium with or without MKs from the BM of TGFβ1 MKΔ/Δ and TGFβ1 fl/fl mice after 14 days. Representative alkaline phosphatase staining images and quantification of the activity of alkaline phosphatase was calculated ( n = 6 per group). g LepR + SSCs were induced in osteogenic differentiation medium with or without MKs from the BM of TGFβ1 MKΔ/Δ and TGFβ1 fl/fl mice after 21 days. Representative alizarin red staining images and quantification of matrix mineralization was calculated ( n = 6 per group). Data on graphs are shown as mean ± SD. One-way ANOVA was used to analyze the data in a – e and an unpaired two-tailed t -test was used to analyze the data in f and g . * P < 0.05, ** P < 0.01 and *** P < 0.001. For all panels in this figure, data are representative of three independent experiments.

    Article Snippet: Briefly, the bone sections were incubated with primary antibodies against mouse osteocalcin (A20800, ABclonal), Ki67 (AF7617, R&D), PTP1B (bs-55182R, Bioss), Slc39a14 (A10413, ABclonal), leptin receptor (bs-0410R, Bioss), CHOP ( A21902 , ABclonal), F4/80 (30325, CST), TGFβ1 (ab313729, abcam), vWF (bsm-52775R, Bioss), osterix (ab209484, abcam) and Smad2 (A7699, ABclonal) overnight at 4 °C and incubated with secondary antibodies for 1 h at 37 °C.

    Techniques: Micro-CT, Staining, Activity Assay, Expressing, Two Tailed Test

    a RNA-seq analysis revealed changes in gene expression in LepR + SSCs co-cultured with MKs ( n = 3 each). b qPCR analysis of the expressions of S mad2 and Slc39a14 in LepR + SSCs, with or without MKs, from WT mice ( n = 6 per group). c Western blotting analysis of the expression of Smad2 and Slc39a14 in LepR + SSCs, with or without MKs, from WT mice ( n = 3 per group). d Representative immunostaining images of Smad2 (red) and Slc39a14 (green) in LepR + SSCs, with or without MKs, from the BM of TGFβ1 MKΔ/Δ and TGFβ1 fl/fl mice ( n = 6 per group). Scale bar, 100 µm. e Colocalization of Smad2 (red) with Slc39a14 (green) in LepR + SSCs, with or without MKs, from the BM of TGFβ1 MKΔ/Δ and TGFβ1 fl/fl mice ( n = 6 per group). Ctrl, control. f A schematic representation of the neural network model of Smad2 binding to the promoter region of Slc39a14, predicted by AlphaFold 3. g A plot of the predicted aligned error of the complex predicted by AlphaFold 3 (pTM + ipTM = 0.91). h A plot of the binding site and amino acid residues of Smad2–Slc39a14 analyzed by PyMol. i Dual-luciferase assays of 293T cotransfected with WT or mutated Slc39a14 (LUC), combined with pcDNA3.1-Smad2 or pcDNA3.1 vetor. j ChIP assay of Smad2 binding to Slc39a14 promoters in LepR + SSCs transfected with pcDNA3.1-Smad2 or pcDNA3.1. Immunoprecipitated DNA and the input DNA were detected by PCR. Primer sequences were designed for Slc39a14 promoter regions located in the promoter region of the Slc39a14 gene, with IgG as a negative control. Data on graphs are shown as mean ± SD. An unpaired two-tailed t -test was used to analyze the data in b and i . * P < 0.05, ** P < 0.01 and *** P < 0.001. For all panels in this figure, data are representative of three independent experiments.

    Journal: Experimental & Molecular Medicine

    Article Title: Megakaryocytic TGFβ1 orchestrates osteogenesis of LepR + SSCs to alleviate radiation-induced bone loss

    doi: 10.1038/s12276-025-01612-z

    Figure Lengend Snippet: a RNA-seq analysis revealed changes in gene expression in LepR + SSCs co-cultured with MKs ( n = 3 each). b qPCR analysis of the expressions of S mad2 and Slc39a14 in LepR + SSCs, with or without MKs, from WT mice ( n = 6 per group). c Western blotting analysis of the expression of Smad2 and Slc39a14 in LepR + SSCs, with or without MKs, from WT mice ( n = 3 per group). d Representative immunostaining images of Smad2 (red) and Slc39a14 (green) in LepR + SSCs, with or without MKs, from the BM of TGFβ1 MKΔ/Δ and TGFβ1 fl/fl mice ( n = 6 per group). Scale bar, 100 µm. e Colocalization of Smad2 (red) with Slc39a14 (green) in LepR + SSCs, with or without MKs, from the BM of TGFβ1 MKΔ/Δ and TGFβ1 fl/fl mice ( n = 6 per group). Ctrl, control. f A schematic representation of the neural network model of Smad2 binding to the promoter region of Slc39a14, predicted by AlphaFold 3. g A plot of the predicted aligned error of the complex predicted by AlphaFold 3 (pTM + ipTM = 0.91). h A plot of the binding site and amino acid residues of Smad2–Slc39a14 analyzed by PyMol. i Dual-luciferase assays of 293T cotransfected with WT or mutated Slc39a14 (LUC), combined with pcDNA3.1-Smad2 or pcDNA3.1 vetor. j ChIP assay of Smad2 binding to Slc39a14 promoters in LepR + SSCs transfected with pcDNA3.1-Smad2 or pcDNA3.1. Immunoprecipitated DNA and the input DNA were detected by PCR. Primer sequences were designed for Slc39a14 promoter regions located in the promoter region of the Slc39a14 gene, with IgG as a negative control. Data on graphs are shown as mean ± SD. An unpaired two-tailed t -test was used to analyze the data in b and i . * P < 0.05, ** P < 0.01 and *** P < 0.001. For all panels in this figure, data are representative of three independent experiments.

    Article Snippet: Briefly, the bone sections were incubated with primary antibodies against mouse osteocalcin (A20800, ABclonal), Ki67 (AF7617, R&D), PTP1B (bs-55182R, Bioss), Slc39a14 (A10413, ABclonal), leptin receptor (bs-0410R, Bioss), CHOP ( A21902 , ABclonal), F4/80 (30325, CST), TGFβ1 (ab313729, abcam), vWF (bsm-52775R, Bioss), osterix (ab209484, abcam) and Smad2 (A7699, ABclonal) overnight at 4 °C and incubated with secondary antibodies for 1 h at 37 °C.

    Techniques: RNA Sequencing, Gene Expression, Cell Culture, Western Blot, Expressing, Immunostaining, Control, Binding Assay, Luciferase, Transfection, Immunoprecipitation, Negative Control, Two Tailed Test

    a Serum zinc concentration in mice 4 weeks after irradiation with administration of TPO or vehicle ( n = 6 per group). b Representative fluozin-3 images and quantitative analysis of LepR + SSCs, with or without MKs, after irradiation ( n = 6 per group). c Representative immunostaining images of Slc39a14 (green) in LepR + SSCs, with or without MKs, after irradiation ( n = 6 per group). Scale bar, 100 µm. d KEGG enrichment analysis of upregulated pathways in LepR + SSCs after irradiation. e GO enrichment analysis of downregulated functions in LepR + SSCs after co-culture with MKs. The top ten enriched GO terms ( P < 0.05) are shown. f Western blotting analysis of the expression of Slc39a14, PTP1B, p-eIF2α, ATF4 and CHOP in LepR + SSCs after co-culture with MKs ( n = 3 per group). g Representative immunostaining images of CHOP (green) in LepR + SSCs, with or without MKs, after irradiation ( n = 6 per group). Scale bar, 100 µm. h Transmission electron microscopy images of LepR + SSCs after irradiation co-culture with MKs ( n = 3 per group). Data on graphs are shown as mean ± SD. One-way ANOVA was used to analyze the data in a – d and g . * P < 0.05, ** P < 0.01 and *** P < 0.001. For all panels in this figure, data are representative of three independent experiments.

    Journal: Experimental & Molecular Medicine

    Article Title: Megakaryocytic TGFβ1 orchestrates osteogenesis of LepR + SSCs to alleviate radiation-induced bone loss

    doi: 10.1038/s12276-025-01612-z

    Figure Lengend Snippet: a Serum zinc concentration in mice 4 weeks after irradiation with administration of TPO or vehicle ( n = 6 per group). b Representative fluozin-3 images and quantitative analysis of LepR + SSCs, with or without MKs, after irradiation ( n = 6 per group). c Representative immunostaining images of Slc39a14 (green) in LepR + SSCs, with or without MKs, after irradiation ( n = 6 per group). Scale bar, 100 µm. d KEGG enrichment analysis of upregulated pathways in LepR + SSCs after irradiation. e GO enrichment analysis of downregulated functions in LepR + SSCs after co-culture with MKs. The top ten enriched GO terms ( P < 0.05) are shown. f Western blotting analysis of the expression of Slc39a14, PTP1B, p-eIF2α, ATF4 and CHOP in LepR + SSCs after co-culture with MKs ( n = 3 per group). g Representative immunostaining images of CHOP (green) in LepR + SSCs, with or without MKs, after irradiation ( n = 6 per group). Scale bar, 100 µm. h Transmission electron microscopy images of LepR + SSCs after irradiation co-culture with MKs ( n = 3 per group). Data on graphs are shown as mean ± SD. One-way ANOVA was used to analyze the data in a – d and g . * P < 0.05, ** P < 0.01 and *** P < 0.001. For all panels in this figure, data are representative of three independent experiments.

    Article Snippet: Briefly, the bone sections were incubated with primary antibodies against mouse osteocalcin (A20800, ABclonal), Ki67 (AF7617, R&D), PTP1B (bs-55182R, Bioss), Slc39a14 (A10413, ABclonal), leptin receptor (bs-0410R, Bioss), CHOP ( A21902 , ABclonal), F4/80 (30325, CST), TGFβ1 (ab313729, abcam), vWF (bsm-52775R, Bioss), osterix (ab209484, abcam) and Smad2 (A7699, ABclonal) overnight at 4 °C and incubated with secondary antibodies for 1 h at 37 °C.

    Techniques: Concentration Assay, Irradiation, Immunostaining, Co-Culture Assay, Western Blot, Expressing, Transmission Assay, Electron Microscopy

    a Representative immunostaining images of LepR (red) and PTP1B (green) in the BM of irradiated mice ( n = 6 mice per group). Scale bar, 100 µm. b Representative immunostaining images of LepR (red) and PTP1B (green) in the BM of MK deleted mice and their littermate controls after irradiation ( n = 6 mice per group). Scale bar, 100 µm. c Representative immunostaining images of PTP1B in LepR + cells, with or without, MKs from the BM of TGFβ1 MKΔ/Δ and TGFβ1 fl/fl mice after irradiation ( n = 6 per group). Scale bar, 100 µm. d Western blotting analysis of the expression of PTP1B and p-Stat3 in LepR + SSCs after co-culture with MKs ( n = 3 per group), inh = inhibitor. e Western blotting analysis of the expression of PTP1B and p-Stat3 in LepR + SSCs after co-culture with MKs from the BM of TGFβ1 MKΔ/Δ and TGFβ1 fl/fl mice ( n = 3 per group). Data on graphs are shown as mean ± SD. One-way ANOVA was used to analyze the data in c and d . * P < 0.05, ** P < 0.01 and *** P < 0.001. For all panels in this figure, data are representative of three independent experiments.

    Journal: Experimental & Molecular Medicine

    Article Title: Megakaryocytic TGFβ1 orchestrates osteogenesis of LepR + SSCs to alleviate radiation-induced bone loss

    doi: 10.1038/s12276-025-01612-z

    Figure Lengend Snippet: a Representative immunostaining images of LepR (red) and PTP1B (green) in the BM of irradiated mice ( n = 6 mice per group). Scale bar, 100 µm. b Representative immunostaining images of LepR (red) and PTP1B (green) in the BM of MK deleted mice and their littermate controls after irradiation ( n = 6 mice per group). Scale bar, 100 µm. c Representative immunostaining images of PTP1B in LepR + cells, with or without, MKs from the BM of TGFβ1 MKΔ/Δ and TGFβ1 fl/fl mice after irradiation ( n = 6 per group). Scale bar, 100 µm. d Western blotting analysis of the expression of PTP1B and p-Stat3 in LepR + SSCs after co-culture with MKs ( n = 3 per group), inh = inhibitor. e Western blotting analysis of the expression of PTP1B and p-Stat3 in LepR + SSCs after co-culture with MKs from the BM of TGFβ1 MKΔ/Δ and TGFβ1 fl/fl mice ( n = 3 per group). Data on graphs are shown as mean ± SD. One-way ANOVA was used to analyze the data in c and d . * P < 0.05, ** P < 0.01 and *** P < 0.001. For all panels in this figure, data are representative of three independent experiments.

    Article Snippet: Briefly, the bone sections were incubated with primary antibodies against mouse osteocalcin (A20800, ABclonal), Ki67 (AF7617, R&D), PTP1B (bs-55182R, Bioss), Slc39a14 (A10413, ABclonal), leptin receptor (bs-0410R, Bioss), CHOP ( A21902 , ABclonal), F4/80 (30325, CST), TGFβ1 (ab313729, abcam), vWF (bsm-52775R, Bioss), osterix (ab209484, abcam) and Smad2 (A7699, ABclonal) overnight at 4 °C and incubated with secondary antibodies for 1 h at 37 °C.

    Techniques: Immunostaining, Irradiation, Western Blot, Expressing, Co-Culture Assay

    a Representative micro-CT images of longitudinal section femurs, cross-sectional view of the distal femurs and reconstructed trabecular structure of the region of interest from mice injected with TPO or vehicle after irradiation ( n = 6 mice per group). b Quantitative micro-CT analysis of the TB fraction (BV/TV, Tb.N, Tb.Th, Tb.Sp, BMD and Ct.Th) in mice injected with TPO or vehicle after irradiation ( n = 6 mice per group). c Representative immunostaining images of OCN (red) in the TB and EB of the mice injected with TPO or vehicle after irradiation. The quantification of OCN cells is shown on the right ( n = 6 mice per group). Scale bar, 100 µm. d HE staining demonstrating B.Ar/T.Ar (bone area/total area) and the presence of MKs in the osteogenic niche of control or irradiated mice 8 weeks after injection with TPO ( n = 6 mice per group). Scale bar, 100 µm. e Von Kossa staining showing mineralization of bone matrix in control or irradiated mice 8 weeks after injected with TPO ( n = 6 mice per group). Scale bar, 1 mm. f Representative immunostaining images of perilipin (red) and osteopontin (OPN, green) in the BM of irradiation mice ( n = 6 mice per group). Scale bar, 100 µm. g Colocalization of LepR (red) with Slc39a14 (green) in the BM of irradiation mice ( n = 6 mice per group). Scale bar, 100 µm. Data on graphs are shown as mean ± SD. One-way ANOVA was used to analyze the data in b – g . * P < 0.05, ** P < 0.01 and *** P < 0.001. For all panels in this figure, data are representative of three independent experiments.

    Journal: Experimental & Molecular Medicine

    Article Title: Megakaryocytic TGFβ1 orchestrates osteogenesis of LepR + SSCs to alleviate radiation-induced bone loss

    doi: 10.1038/s12276-025-01612-z

    Figure Lengend Snippet: a Representative micro-CT images of longitudinal section femurs, cross-sectional view of the distal femurs and reconstructed trabecular structure of the region of interest from mice injected with TPO or vehicle after irradiation ( n = 6 mice per group). b Quantitative micro-CT analysis of the TB fraction (BV/TV, Tb.N, Tb.Th, Tb.Sp, BMD and Ct.Th) in mice injected with TPO or vehicle after irradiation ( n = 6 mice per group). c Representative immunostaining images of OCN (red) in the TB and EB of the mice injected with TPO or vehicle after irradiation. The quantification of OCN cells is shown on the right ( n = 6 mice per group). Scale bar, 100 µm. d HE staining demonstrating B.Ar/T.Ar (bone area/total area) and the presence of MKs in the osteogenic niche of control or irradiated mice 8 weeks after injection with TPO ( n = 6 mice per group). Scale bar, 100 µm. e Von Kossa staining showing mineralization of bone matrix in control or irradiated mice 8 weeks after injected with TPO ( n = 6 mice per group). Scale bar, 1 mm. f Representative immunostaining images of perilipin (red) and osteopontin (OPN, green) in the BM of irradiation mice ( n = 6 mice per group). Scale bar, 100 µm. g Colocalization of LepR (red) with Slc39a14 (green) in the BM of irradiation mice ( n = 6 mice per group). Scale bar, 100 µm. Data on graphs are shown as mean ± SD. One-way ANOVA was used to analyze the data in b – g . * P < 0.05, ** P < 0.01 and *** P < 0.001. For all panels in this figure, data are representative of three independent experiments.

    Article Snippet: Briefly, the bone sections were incubated with primary antibodies against mouse osteocalcin (A20800, ABclonal), Ki67 (AF7617, R&D), PTP1B (bs-55182R, Bioss), Slc39a14 (A10413, ABclonal), leptin receptor (bs-0410R, Bioss), CHOP ( A21902 , ABclonal), F4/80 (30325, CST), TGFβ1 (ab313729, abcam), vWF (bsm-52775R, Bioss), osterix (ab209484, abcam) and Smad2 (A7699, ABclonal) overnight at 4 °C and incubated with secondary antibodies for 1 h at 37 °C.

    Techniques: Micro-CT, Injection, Irradiation, Immunostaining, Staining, Control

    a Representative micro-CT images of longitudinal section femurs, cross-sectional view of the distal femurs and reconstructed trabecular structure of the region of interest from Slc39a14 leprΔ/Δ mice and their littermate controls (Slc39a14 fl/fl mice) ( n = 6 mice per group). b Quantitative micro-CT analysis of the TB fraction (BV/TV, Tb.N, Tb.Th, Tb.Sp, BMD and Ct.Th) in Slc39a14 leprΔ/Δ mice and their littermate controls (Slc39a14 fl/fl mice) ( n = 6 mice per group). c Quantitative biomechanical analysis of femora (peak load and stiffness) from MK deleted mice and their littermate controls (Slc39a14 fl/fl mice) ( n = 6 mice per group). d Serum zinc concentration in Slc39a14 leprΔ/Δ mice and their littermate controls (Slc39a14 fl/fl mice) ( n = 6 per group). e Von Kossa staining showing mineralization of bone matrix in Slc39a14 leprΔ/Δ mice and their littermate controls (Slc39a14 fl/fl mice) ( n = 6 per group). f Representative micro-CT images of longitudinal section femurs, cross-sectional view of the distal femurs and reconstructed trabecular structure of the region of interest from Slc39a14 leprΔ/Δ mice injected with TPO or vehicle after irradiation ( n = 6 mice per group). g Representative immunostaining images of OCN (red) in the TB and EB of the Slc39a14 leprΔ/Δ mice injected with TPO or vehicle after irradiation. The quantification of OCN cells is shown on the right ( n = 6 mice per group). Scale bar, 100 µm. h Representative immunostaining images of LepR (red) and PTP1B (green) in the BM of the Slc39a14 leprΔ/Δ mice injected with TPO or vehicle after irradiation. Scale bar, 100 µm. i Colocalization of LepR (red) with Ki67 (green) in the BM of the Slc39a14 leprΔ/Δ mice injected with TPO or vehicle after irradiation ( n = 6 mice per group). Scale bar, 100 µm. j Representative immunostaining images of TUNEL (green) in the BM of the Slc39a14 leprΔ/Δ mice injected with TPO or vehicle after irradiation. The quantification of tunel positive cells is shown on the right ( n = 6 mice per group). Scale bar, 100 µm. Data on graphs are shown as mean ± SD. An unpaired two-tailed t -test was used to analyze the data in b – d , g , i and j . * P < 0.05, ** P < 0.01 and *** P < 0.001. For all panels in this figure, data are representative of three independent experiments.

    Journal: Experimental & Molecular Medicine

    Article Title: Megakaryocytic TGFβ1 orchestrates osteogenesis of LepR + SSCs to alleviate radiation-induced bone loss

    doi: 10.1038/s12276-025-01612-z

    Figure Lengend Snippet: a Representative micro-CT images of longitudinal section femurs, cross-sectional view of the distal femurs and reconstructed trabecular structure of the region of interest from Slc39a14 leprΔ/Δ mice and their littermate controls (Slc39a14 fl/fl mice) ( n = 6 mice per group). b Quantitative micro-CT analysis of the TB fraction (BV/TV, Tb.N, Tb.Th, Tb.Sp, BMD and Ct.Th) in Slc39a14 leprΔ/Δ mice and their littermate controls (Slc39a14 fl/fl mice) ( n = 6 mice per group). c Quantitative biomechanical analysis of femora (peak load and stiffness) from MK deleted mice and their littermate controls (Slc39a14 fl/fl mice) ( n = 6 mice per group). d Serum zinc concentration in Slc39a14 leprΔ/Δ mice and their littermate controls (Slc39a14 fl/fl mice) ( n = 6 per group). e Von Kossa staining showing mineralization of bone matrix in Slc39a14 leprΔ/Δ mice and their littermate controls (Slc39a14 fl/fl mice) ( n = 6 per group). f Representative micro-CT images of longitudinal section femurs, cross-sectional view of the distal femurs and reconstructed trabecular structure of the region of interest from Slc39a14 leprΔ/Δ mice injected with TPO or vehicle after irradiation ( n = 6 mice per group). g Representative immunostaining images of OCN (red) in the TB and EB of the Slc39a14 leprΔ/Δ mice injected with TPO or vehicle after irradiation. The quantification of OCN cells is shown on the right ( n = 6 mice per group). Scale bar, 100 µm. h Representative immunostaining images of LepR (red) and PTP1B (green) in the BM of the Slc39a14 leprΔ/Δ mice injected with TPO or vehicle after irradiation. Scale bar, 100 µm. i Colocalization of LepR (red) with Ki67 (green) in the BM of the Slc39a14 leprΔ/Δ mice injected with TPO or vehicle after irradiation ( n = 6 mice per group). Scale bar, 100 µm. j Representative immunostaining images of TUNEL (green) in the BM of the Slc39a14 leprΔ/Δ mice injected with TPO or vehicle after irradiation. The quantification of tunel positive cells is shown on the right ( n = 6 mice per group). Scale bar, 100 µm. Data on graphs are shown as mean ± SD. An unpaired two-tailed t -test was used to analyze the data in b – d , g , i and j . * P < 0.05, ** P < 0.01 and *** P < 0.001. For all panels in this figure, data are representative of three independent experiments.

    Article Snippet: Briefly, the bone sections were incubated with primary antibodies against mouse osteocalcin (A20800, ABclonal), Ki67 (AF7617, R&D), PTP1B (bs-55182R, Bioss), Slc39a14 (A10413, ABclonal), leptin receptor (bs-0410R, Bioss), CHOP ( A21902 , ABclonal), F4/80 (30325, CST), TGFβ1 (ab313729, abcam), vWF (bsm-52775R, Bioss), osterix (ab209484, abcam) and Smad2 (A7699, ABclonal) overnight at 4 °C and incubated with secondary antibodies for 1 h at 37 °C.

    Techniques: Micro-CT, Concentration Assay, Staining, Injection, Irradiation, Immunostaining, TUNEL Assay, Two Tailed Test

    Age was not associated with any of the brain protein loads in the DS and DSAD groups. In the controls, we found associations between age and LepR ( p = 0.01), pSTAT3‐727 ( p = 0.04), and pSTAT3‐705 ( p = 0.002). DS, Down syndrome; DSAD, Down syndrome and Alzheimer's disease; LepR, leptin receptor; pSTAT3, phosphorylated STAT3.

    Journal: Alzheimer's & Dementia

    Article Title: Leptin levels are associated with body mass index and Alzheimer's disease in Down syndrome

    doi: 10.1002/alz.70448

    Figure Lengend Snippet: Age was not associated with any of the brain protein loads in the DS and DSAD groups. In the controls, we found associations between age and LepR ( p = 0.01), pSTAT3‐727 ( p = 0.04), and pSTAT3‐705 ( p = 0.002). DS, Down syndrome; DSAD, Down syndrome and Alzheimer's disease; LepR, leptin receptor; pSTAT3, phosphorylated STAT3.

    Article Snippet: Sections were incubated overnight at 4°C in the primary antibody: leptin (1:600; bs‐0108R, Bioss, USA), LepR (1:600; bs‐0109R, Bioss, USA), pSTAT3‐727 (1:600; 44‐384G, ThermoFisher Scientific, USA), pSTAT3‐705 (1:2500; PA5‐121259, ThermoFisher Scientific, USA), and SOCS3 (1:400; bs‐0580R, Bioss, USA).

    Techniques:

    List of antibodies used in the study (IF—immunofluorescence staining; WB—Western blot analysis).

    Journal: International Journal of Molecular Sciences

    Article Title: Chemerin Stimulates the Secretory Activity of BME-UV1 Bovine Mammary Epithelial Cells

    doi: 10.3390/ijms25084147

    Figure Lengend Snippet: List of antibodies used in the study (IF—immunofluorescence staining; WB—Western blot analysis).

    Article Snippet: Leptin receptor Rabbit Polyclonal Antibody , Bioss Antibodies , bs-0961R , 1:200 , IF.

    Techniques: Staining

    Viability of BME-UV1 bovine mammary epithelial cells treated for 24 h with different concentrations of chemerin, leptin, or adiponectin. Graphs present cell viability measured using the MTT assay. The viability of untreated control cells (0) was designated as 100%. Results are presented as means ± standard deviation of three independent experiments.

    Journal: International Journal of Molecular Sciences

    Article Title: Chemerin Stimulates the Secretory Activity of BME-UV1 Bovine Mammary Epithelial Cells

    doi: 10.3390/ijms25084147

    Figure Lengend Snippet: Viability of BME-UV1 bovine mammary epithelial cells treated for 24 h with different concentrations of chemerin, leptin, or adiponectin. Graphs present cell viability measured using the MTT assay. The viability of untreated control cells (0) was designated as 100%. Results are presented as means ± standard deviation of three independent experiments.

    Article Snippet: Leptin receptor Rabbit Polyclonal Antibody , Bioss Antibodies , bs-0961R , 1:200 , IF.

    Techniques: MTT Assay, Control, Standard Deviation

    Immunofluorescence staining of leptin receptor in BME-UV1 bovine mammary epithelial cells. Panels I and II present the localization of leptin receptor detected using primary antibodies (cat. no. bs-0961; Bioss Antibodies), followed by secondary antibodies conjugated with Alexa Fluor 488 dye (green fluorescence); nuclei were counterstained with 7-amino actinomycin (7-AAD, red fluorescence). Panel III presents no primary antibody control. Micrographs were taken at 600× magnification. Scale bar: 20 μm.

    Journal: International Journal of Molecular Sciences

    Article Title: Chemerin Stimulates the Secretory Activity of BME-UV1 Bovine Mammary Epithelial Cells

    doi: 10.3390/ijms25084147

    Figure Lengend Snippet: Immunofluorescence staining of leptin receptor in BME-UV1 bovine mammary epithelial cells. Panels I and II present the localization of leptin receptor detected using primary antibodies (cat. no. bs-0961; Bioss Antibodies), followed by secondary antibodies conjugated with Alexa Fluor 488 dye (green fluorescence); nuclei were counterstained with 7-amino actinomycin (7-AAD, red fluorescence). Panel III presents no primary antibody control. Micrographs were taken at 600× magnification. Scale bar: 20 μm.

    Article Snippet: Leptin receptor Rabbit Polyclonal Antibody , Bioss Antibodies , bs-0961R , 1:200 , IF.

    Techniques: Immunofluorescence, Staining, Fluorescence, Control

    Evaluation of apoptosis in BME-UV1 bovine mammary epithelial cells treated with chemerin (Ch10 = 10 ng/mL, Ch100 = 100 ng/mL), leptin (L10 = 10 ng/mL, L100 = 100 ng/mL), or adiponectin (A10 = 10 ng/mL, A500 = 500 ng/mL) for 24 h. ( A ) Representative dot-plots of Annexin V/PI double staining in untreated control cells; ( B ) graph showing percentage of apoptotic cells (sum of Annexin V pos /PI neg and Annexin V pos /Pi pos cells) in control and adipokine-treated cells; ( C ) representative images of Western blot (WB) analysis of apoptotic markers cleaved caspase 3 and bax, where gapdh was used as a reference protein; ( D , E ) graphs showing the results of the densitometric analysis of WB images; the integrated optical density (IOD) of cleaved caspase 3 and bax bands was normalized to the IOD of gapdh bands. Results are presented as means ± standard deviation of three or four independent experiments.

    Journal: International Journal of Molecular Sciences

    Article Title: Chemerin Stimulates the Secretory Activity of BME-UV1 Bovine Mammary Epithelial Cells

    doi: 10.3390/ijms25084147

    Figure Lengend Snippet: Evaluation of apoptosis in BME-UV1 bovine mammary epithelial cells treated with chemerin (Ch10 = 10 ng/mL, Ch100 = 100 ng/mL), leptin (L10 = 10 ng/mL, L100 = 100 ng/mL), or adiponectin (A10 = 10 ng/mL, A500 = 500 ng/mL) for 24 h. ( A ) Representative dot-plots of Annexin V/PI double staining in untreated control cells; ( B ) graph showing percentage of apoptotic cells (sum of Annexin V pos /PI neg and Annexin V pos /Pi pos cells) in control and adipokine-treated cells; ( C ) representative images of Western blot (WB) analysis of apoptotic markers cleaved caspase 3 and bax, where gapdh was used as a reference protein; ( D , E ) graphs showing the results of the densitometric analysis of WB images; the integrated optical density (IOD) of cleaved caspase 3 and bax bands was normalized to the IOD of gapdh bands. Results are presented as means ± standard deviation of three or four independent experiments.

    Article Snippet: Leptin receptor Rabbit Polyclonal Antibody , Bioss Antibodies , bs-0961R , 1:200 , IF.

    Techniques: Double Staining, Control, Western Blot, Standard Deviation

    Concentration of αS1-casein in BME-UV1 cells ( A ) and media ( B ) collected after 24 h of culture. Cell were treated with chemerin (Ch10 = 10 ng/mL, Ch100 = 100 ng/mL), leptin (L10 = 10 ng/mL, L100 = 100 ng/mL) or adiponectin (A10 = 10 ng/mL, A500 = 500 ng/mL). Untreated cells were used as a negative control, whereas cells exposed for 24 h to the lactogenic hormone prolactin (PRL, 1 µg/mL) were used as a positive control. Results are presented as means ± standard deviation of three independent experiments. Means followed by a common letter are not significantly different according to one-way ANOVA with Tukey’s multiple comparison post-test, at the 5% level of significance ( p < 0.05).

    Journal: International Journal of Molecular Sciences

    Article Title: Chemerin Stimulates the Secretory Activity of BME-UV1 Bovine Mammary Epithelial Cells

    doi: 10.3390/ijms25084147

    Figure Lengend Snippet: Concentration of αS1-casein in BME-UV1 cells ( A ) and media ( B ) collected after 24 h of culture. Cell were treated with chemerin (Ch10 = 10 ng/mL, Ch100 = 100 ng/mL), leptin (L10 = 10 ng/mL, L100 = 100 ng/mL) or adiponectin (A10 = 10 ng/mL, A500 = 500 ng/mL). Untreated cells were used as a negative control, whereas cells exposed for 24 h to the lactogenic hormone prolactin (PRL, 1 µg/mL) were used as a positive control. Results are presented as means ± standard deviation of three independent experiments. Means followed by a common letter are not significantly different according to one-way ANOVA with Tukey’s multiple comparison post-test, at the 5% level of significance ( p < 0.05).

    Article Snippet: Leptin receptor Rabbit Polyclonal Antibody , Bioss Antibodies , bs-0961R , 1:200 , IF.

    Techniques: Concentration Assay, Negative Control, Positive Control, Standard Deviation, Comparison

    Panels of confocal micrographs presenting the immunofluorescence staining of BME-UV1 cells cultured on Matrigel for 11 days in control growth medium (Ctrl.) or medium supplemented with chemerin (Ch10 = 10 ng/mL, Ch100 = 100 ng/mL), leptin (L10 = 10 ng/mL, L100 = 100 ng/mL), or adiponectin (A10 = 10 ng/mL, A500 = 500 ng/mL). Cells were stained with primary antibodies against β1-integrin and secondary antibodies conjugated with Alexa Fluor 488 dye (green fluorescence); Alexa Fluor 594 phalloidin, detecting F-actin (red fluorescence), and nuclei counterstained with Hoechst 33342 (blue fluorescence). Images were taken at 600× magnification and are representative for three independent experiments. Scale bar: 20 μm.

    Journal: International Journal of Molecular Sciences

    Article Title: Chemerin Stimulates the Secretory Activity of BME-UV1 Bovine Mammary Epithelial Cells

    doi: 10.3390/ijms25084147

    Figure Lengend Snippet: Panels of confocal micrographs presenting the immunofluorescence staining of BME-UV1 cells cultured on Matrigel for 11 days in control growth medium (Ctrl.) or medium supplemented with chemerin (Ch10 = 10 ng/mL, Ch100 = 100 ng/mL), leptin (L10 = 10 ng/mL, L100 = 100 ng/mL), or adiponectin (A10 = 10 ng/mL, A500 = 500 ng/mL). Cells were stained with primary antibodies against β1-integrin and secondary antibodies conjugated with Alexa Fluor 488 dye (green fluorescence); Alexa Fluor 594 phalloidin, detecting F-actin (red fluorescence), and nuclei counterstained with Hoechst 33342 (blue fluorescence). Images were taken at 600× magnification and are representative for three independent experiments. Scale bar: 20 μm.

    Article Snippet: Leptin receptor Rabbit Polyclonal Antibody , Bioss Antibodies , bs-0961R , 1:200 , IF.

    Techniques: Immunofluorescence, Staining, Cell Culture, Control, Fluorescence

    Diameters of 3D mammospheres formed by BME-UV1 cells cultured on Matrigel for 11 days. Cells were cultured in control growth medium (Ctrl.) or medium supplemented with chemerin (Ch10 = 10 ng/mL, Ch100 = 100 ng/mL), leptin (L10 = 10 ng/mL, L100 = 100 ng/mL), or adiponectin (A10 = 10 ng/mL, A500 = 500 ng/mL). The diameters of cross sections of the spheroids were measured using ImageJ software ( https://ij.imjoy.io accessed on 2 April 2024). Results are presented as means ± standard deviation of at least 14 spheroids per experimental condition photographed using a confocal microscope equipped with a digital camera.

    Journal: International Journal of Molecular Sciences

    Article Title: Chemerin Stimulates the Secretory Activity of BME-UV1 Bovine Mammary Epithelial Cells

    doi: 10.3390/ijms25084147

    Figure Lengend Snippet: Diameters of 3D mammospheres formed by BME-UV1 cells cultured on Matrigel for 11 days. Cells were cultured in control growth medium (Ctrl.) or medium supplemented with chemerin (Ch10 = 10 ng/mL, Ch100 = 100 ng/mL), leptin (L10 = 10 ng/mL, L100 = 100 ng/mL), or adiponectin (A10 = 10 ng/mL, A500 = 500 ng/mL). The diameters of cross sections of the spheroids were measured using ImageJ software ( https://ij.imjoy.io accessed on 2 April 2024). Results are presented as means ± standard deviation of at least 14 spheroids per experimental condition photographed using a confocal microscope equipped with a digital camera.

    Article Snippet: Leptin receptor Rabbit Polyclonal Antibody , Bioss Antibodies , bs-0961R , 1:200 , IF.

    Techniques: Cell Culture, Control, Software, Standard Deviation, Microscopy

    List of antibodies used in the study (IF—immunofluorescence staining; WB—Western blot analysis).

    Journal: International Journal of Molecular Sciences

    Article Title: Chemerin Stimulates the Secretory Activity of BME-UV1 Bovine Mammary Epithelial Cells

    doi: 10.3390/ijms25084147

    Figure Lengend Snippet: List of antibodies used in the study (IF—immunofluorescence staining; WB—Western blot analysis).

    Article Snippet: Leptin receptor Rabbit Polyclonal Antibody , Bioss Antibodies , bs-0961R , 1:200 , IF.

    Techniques: Staining