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

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

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

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

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

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

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

Journal: Cancer Research and Treatment : Official Journal of Korean Cancer Association

Article Title: Synergistic Activation of LEPR and ADRB2 Induced by Leptin Enhances Reactive Oxygen Specie Generation in Triple-Negative Breast Cancer Cells

doi: 10.4143/crt.2024.368

Figure Lengend Snippet: Leptin upregulates ADRB2 expression in TNBC cells. (A) Quantitative PCR was utilized to assess the expression levels of ADRB2 in 4T1 (n=3) and EMT6 (n=6) cells following stimulation with 50 ng/mL or 100 ng/mL leptin. (B) Western blotting was utilized to assess the expression levels of ADRB2 in 4T1 and EMT6 cells following stimulation with 50 ng/mL or 100 ng/mL leptin. n=4. (C, D) Quantitative PCR (C) and western blotting (D) were employed to analyze the expression of ADRB2 in 4T1 and EMT6 cells treated with leptin (100 ng/mL) and/or Allo (100 nM). n=3. (E) Representative images of immunofluorescence double staining showing the expression and co-localization of LEPR (Alexa Fluor 488 Conjugated) and ADRB2 (Alexa Fluor 594-conjugated) in the presence of leptin and/or Allo, or the ADRB2 inhibitor ICI118551 (ICI). (F-J) Statistical curves depicting the fluorescence intensity of LEPR and ADRB2 in the co-localization images for different experimental groups. (K) Quantitative analysis of LEPR-ADRB2 co-localization on cell membranes in each group, evaluated using Pearson’s correlation coefficient. Data are presented as mean±SEM, n=3. ADRB2, adrenergic receptor β2; LEPR, leptin receptor; PCR, polymerase chain reaction; SEM, standard error of mean; TNBC, triple-negative breast cancer. All data are presented as the mean±SEM. ns, no significance; * p < 0.05, ** p < 0.01.

Article Snippet: The cells were blocked with 5% goat serum for 30 minutes and then incubated overnight at 4°C with a 1:100 dilution of anti-LEPR antibody (sc-8391, Santa Cruz Biotechnology) and a 1:200 dilution of anti-ADRB2 antibody (ab128136, Abcam).

Techniques: Expressing, Real-time Polymerase Chain Reaction, Western Blot, Immunofluorescence, Double Staining, Fluorescence, Polymerase Chain Reaction

Journal: Cancer Research and Treatment : Official Journal of Korean Cancer Association

Article Title: Synergistic Activation of LEPR and ADRB2 Induced by Leptin Enhances Reactive Oxygen Specie Generation in Triple-Negative Breast Cancer Cells

doi: 10.4143/crt.2024.368

Figure Lengend Snippet: LEPR and ADRB2 in leptin-induced breast cancer progression in vivo . (A) Female BALB/c mice were subcutaneously inoculated with 1×10 5 4T1 cells into the mammary fat pad to establish the breast tumor animal model. Drug administration commenced 7 days postinoculation and continued for 21 days. Tumors, blood samples, and lungs were collected for analysis. (B) Representative image of tumors and tumor growth curves in different experimental groups. n=5. (C) Tumor weights were measured on day 28. n=5. (D, E) Food intake (D) and body weight curves (E) in different groups. n=5. (F) Metastatic lung nodules were visualized using H&E staining in different groups. (G) GSH content was assessed in both serum and tumor samples collected from different groups. n=3-5. (H, I) Real-time qPCR (H) and western blotting (I) were used to analyze the expression of LEPR, ADRB2, and NOX4 in tumors from different groups. n=3. ADRB2, adrenergic receptor β2; GSH, glutathione; LEPR, leptin receptor; NOX4, NADPH oxidase 4; qPCR, quantitative real-time polymerase chain reacion; SEM, standard error of mean. All data are presented as the mean±SEM. ns, no significance; * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: The cells were blocked with 5% goat serum for 30 minutes and then incubated overnight at 4°C with a 1:100 dilution of anti-LEPR antibody (sc-8391, Santa Cruz Biotechnology) and a 1:200 dilution of anti-ADRB2 antibody (ab128136, Abcam).

Techniques: In Vivo, Animal Model, Staining, Western Blot, Expressing

Journal: Cancer Research and Treatment : Official Journal of Korean Cancer Association

Article Title: Synergistic Activation of LEPR and ADRB2 Induced by Leptin Enhances Reactive Oxygen Specie Generation in Triple-Negative Breast Cancer Cells

doi: 10.4143/crt.2024.368

Figure Lengend Snippet: The clinical significance of LEPR and ADRB2. (A, B) Receiver operating characteristic curve analysis was conducted to evaluate the prognostic significance of LEPR (A) and ADRB2 (B) in breast cancer patients. p < 0.01. (C-E) The correlation between LEPR, ADRB2, NOX4, and GPX4 was analyzed using the bc-GenExMiner database (C) and GEPIA database (D, E). (F) Kaplan-Meier plotter was utilized to illustrate RFS in TNBC patients with high expression of LEPR and ADRB2 combined. p=0.0071. (G) The schematic diagram of the study work. ADRB2, adrenergic receptor β2; AUC, area under the curve; GPX4, glutathione peroxidase 4; HR, hazard ratio; LEPR, leptin receptor; mtROS, mitochondrial reactive oxygen species; NOX4, NADPH oxidase 4; RFS, relapse-free survival; TNBC, triple-negative breast cancer; TNR, true-negative ratio; TPM, transcripts per million; TPR, true-positive ratio.

Article Snippet: The cells were blocked with 5% goat serum for 30 minutes and then incubated overnight at 4°C with a 1:100 dilution of anti-LEPR antibody (sc-8391, Santa Cruz Biotechnology) and a 1:200 dilution of anti-ADRB2 antibody (ab128136, Abcam).

Techniques: Expressing