Journal: Current Pharmaceutical Biotechnology

Article Title: Evaluation of Changes in the Expression Pattern of EDIL3 in Different Grades of Endometrial Cancer

doi: 10.2174/1389201020666190408112822

Figure Lengend Snippet: Optical density of the reaction product for EDIL3 in different grades of endometrial cancer and control.

Article Snippet: Rabbit polyclonal anti-EDIL3 antibody (Novus Biological, USA) was used to perform immunohistochemical staining of the prepared slides.

Techniques: Control

Journal: Cancers

Article Title: Proteomic Analyses of Fibroblast- and Serum-Derived Exosomes Identify QSOX1 as a Marker for Non-invasive Detection of Colorectal Cancer

doi: 10.3390/cancers13061351

Figure Lengend Snippet: Primary fibroblast-derived EXOs display an activation status dependent protein cargo. The established fibroblast cell lines were subjected to cellular protein isolation after 48 h incubation of equal cell numbers in normal growth medium. EXOs were isolated using combined differential centrifugation and ultrafiltration approach following 48 h of fibroblast incubation in starvation medium. Exosomal protein was isolated in biological triplicates and subjected to Mass Spectrometry. Statistical Analysis was performed in Perseus. ( A ) Immunoblot analysis of vimentin (VIM), α-smooth-muscle actin (αSMA), fibroblast activation protein α (FAPα), caveolin 1 (CAV1), cluster of differentiation 90 (CD90)/Thy1 and fibroblast-specific protein 1 (FSP1) in primary fibroblasts, including GAPDH as loading control. ( B ) Particle size distribution in the isolated EXOs (mean of n = 4–10) as measured by nanoparticle tracking analysis (NTA) using ZetaView ® . ( C ) Representative transmission electron microscopy (TEM) image of fibroblast-derived EXOs. ( D ) Immunoblot analysis of EXO markers CD9, CD63, CD81, Flotillin 1 and Tumor susceptibility 101 (TSG101), including calreticulin as negative control. ( E ) Heat map of mass spectrometry data illustrating significantly deregulated vesicular proteins. Proteins with more than 4 undefined values in total or more than 3 undefined values in the NF/CAF subgroups are excluded. Paired t test: q < 0.05, diff. > |1.0|. LFQ: label-free quantification; nda: no data acquired. ( F ) Immunoblot analysis of quiescin sulfhydryl oxidase 1 (QSOX1), actinin α4 (ACTN4), thrombospondin 1 (THBS1) and EGF-like repeats and discoidin domains 3 (EDIL3) in EXOs. ( G ) Immunoblot analysis of QSOX1, ACTN4, THBS1 and EDIL3 in primary fibroblasts whole cell lysate, including GAPDH as loading control. The images of uncropped western blot figures are shown in .

Article Snippet: The following primary antibodies were used: anti-Actin, α-smooth muscle antibody, mouse monoclonal (A5228, Sigma-Aldrich, 1:1000), human EDIL3 antibody (MAB6046, R&D Systems, 1:200), recombinant anti-quiescin Q6 antibody [EPR21866] (ab235444, Abcam, 1:200), mouse (E5Y6Q) mAb IgG2a (61656, Cell Signaling Technology, 1:200) and rabbit (DA1E) mAb IgG XP ® isotype control (3900, Cell Signaling Technology, 1:200).

Techniques: Derivative Assay, Activation Assay, Isolation, Incubation, Centrifugation, Mass Spectrometry, Western Blot, Control, Transmission Assay, Electron Microscopy, Negative Control, Quantitative Proteomics

Journal: Cancers

Article Title: Proteomic Analyses of Fibroblast- and Serum-Derived Exosomes Identify QSOX1 as a Marker for Non-invasive Detection of Colorectal Cancer

doi: 10.3390/cancers13061351

Figure Lengend Snippet: Selected primary fibroblast activation status dependent EXO markers display specificity to blood EXOs in matched CRC patient plasma. ( A ) Particle size distribution of patient-matched plasma EXOs (pEXO) in comparison to whole plasma (wP) and EXO-depleted plasma (edP) as measured by NTA. ( B ) Immunoblot analysis of quiescin sulfhydryl oxidase 1 (QSOX1), thrombospondin 1 (THBS1), EGF-like repeats and discoidin domains 3 (EDIL3), the EXO markers CD9, CD63 and Flotillin 1, including calreticulin as negative control, and albumin in wP, pEXO and edP protein lysates. ( C ) Graphical analysis of immunoblots shown in (B) using ImageJ, comparing signal strength to pEXO mix. Unpaired t -test: * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: The following primary antibodies were used: anti-Actin, α-smooth muscle antibody, mouse monoclonal (A5228, Sigma-Aldrich, 1:1000), human EDIL3 antibody (MAB6046, R&D Systems, 1:200), recombinant anti-quiescin Q6 antibody [EPR21866] (ab235444, Abcam, 1:200), mouse (E5Y6Q) mAb IgG2a (61656, Cell Signaling Technology, 1:200) and rabbit (DA1E) mAb IgG XP ® isotype control (3900, Cell Signaling Technology, 1:200).

Techniques: Activation Assay, Clinical Proteomics, Comparison, Western Blot, Negative Control

Journal: Cancers

Article Title: Proteomic Analyses of Fibroblast- and Serum-Derived Exosomes Identify QSOX1 as a Marker for Non-invasive Detection of Colorectal Cancer

doi: 10.3390/cancers13061351

Figure Lengend Snippet: In vivo marker expression in patient-matched healthy and malignant colon tissue. ( A ) Representative images of paraffin embedded tissue slides of healthy (hC) and malignant (CRC) colon tissue derived from patients 1–3, hematoxylin and eosin (H&E) or immunohistochemically stained for the proteins quiescin sulfhydryl oxidase 1 (QSOX1), thrombospondin 1 (THBS1), EGF-like repeats and discoidin domains 3 (EDIL3), α-smooth-muscle actin (αSMA) and IgG control (Ctrl.). Scale bars equal 250 µm. ( B ) Graphical IHC staining analysis performed in QuPath. From each patient and tissue, a minimum of three representative areas were subjected to graphical and statistical analysis. Mann-Whitney-U test: * p < 0.05, ** p < 0.01. ns = not significant.

Article Snippet: The following primary antibodies were used: anti-Actin, α-smooth muscle antibody, mouse monoclonal (A5228, Sigma-Aldrich, 1:1000), human EDIL3 antibody (MAB6046, R&D Systems, 1:200), recombinant anti-quiescin Q6 antibody [EPR21866] (ab235444, Abcam, 1:200), mouse (E5Y6Q) mAb IgG2a (61656, Cell Signaling Technology, 1:200) and rabbit (DA1E) mAb IgG XP ® isotype control (3900, Cell Signaling Technology, 1:200).

Techniques: In Vivo, Marker, Expressing, Derivative Assay, Staining, Control, Immunohistochemistry, MANN-WHITNEY

Journal: Cancers

Article Title: Proteomic Analyses of Fibroblast- and Serum-Derived Exosomes Identify QSOX1 as a Marker for Non-invasive Detection of Colorectal Cancer

doi: 10.3390/cancers13061351

Figure Lengend Snippet: Exosomal fibroblast activity marker validation in an independent validation cohort. Twenty additional fibroblast cell lines derived from 10 CRC patients were subjected to cellular protein and EXO isolation. Exosomal protein was isolated and subjected to immunoblot. ( A ) Immunoblot analysis of quiescin sulfhydryl oxidase 1 (QSOX1), thrombospondin 1 (THBS1), EGF-like repeats and discoidin domains 3 (EDIL3), in primary fibroblasts-derived EXOs. ( B ) Immunoblot analysis of QSOX1, THBS1 and EDIL3 in primary fibroblasts whole cell lysate, including GAPDH as loading control. ( C ) Graphical analysis of EXO protein immunoblots shown in ( A ) using ImageJ, relative to GAPDH. Mann-Whitney-U test: * p < 0.05. ( D ) Graphical analysis of cellular protein immunoblots shown in ( B ) using ImageJ, relative to GAPDH. Mann-Whitney-U test: * p < 0.05, *** p < 0.001. ns = not significant.

Article Snippet: The following primary antibodies were used: anti-Actin, α-smooth muscle antibody, mouse monoclonal (A5228, Sigma-Aldrich, 1:1000), human EDIL3 antibody (MAB6046, R&D Systems, 1:200), recombinant anti-quiescin Q6 antibody [EPR21866] (ab235444, Abcam, 1:200), mouse (E5Y6Q) mAb IgG2a (61656, Cell Signaling Technology, 1:200) and rabbit (DA1E) mAb IgG XP ® isotype control (3900, Cell Signaling Technology, 1:200).

Techniques: Activity Assay, Marker, Biomarker Discovery, Derivative Assay, Isolation, Western Blot, Control, MANN-WHITNEY

Journal: Science Advances

Article Title: Endogenous DEL-1 restrains melanoma lung metastasis by limiting myeloid cell–associated lung inflammation

doi: 10.1126/sciadv.abc4882

Figure Lengend Snippet: ( A and B ) C57BL/6 mice were treated with intravenous injection of control-Fc or Del-1-Fc 6 hours before an intravenous injection of 2 × 10 5 B16F10 cells expressing DsRed and twice a week treatment thereafter. (A) Representative lung images (top) and the corresponding DsRed fluorescence images (bottom) showing lung metastases ( n = 8 each group) after 14 days of tumor implantation. Photo credit: Hyung-Joon Kwon, University of Ulsan. (B) Quantification of lung metastases by FLI depicted in (A). ( C and D ) Representative (C) and quantitative (D) flow cytometric analysis of different myeloid cells and lymphocytes, graphed on Ly6C by Ly6G and CD3 by NK1.1 dot plots, respectively, in the metastasis-bearing lungs depicted in (A). Horizontal bars indicate the means (D). * P < 0.05.

Article Snippet: To analyze DEL-1 expression by endothelial cells in the metastatic lungs, frozen sections (15 μm) were prepared, permeabilized with 0.1% Triton X-100 in PBS, and incubated with rabbit anti-mouse DEL-1 (1:200; AbFrontier) and Alexa Fluor 594–conjugated anti-mouse CD31 (1:200, MEC13,3; BioLegend) for 12 hours at 4°C, followed by Alexa Fluor 488–conjugated goat anti-rabbit IgG (1:500; Invitrogen) for 1 hour.

Techniques: Injection, Expressing, Fluorescence, Tumor Implantation

Journal: iScience

Article Title: A novel macrolide–Del-1 axis to regenerate bone in old age

doi: 10.1016/j.isci.2024.108798

Figure Lengend Snippet: Local treatment with DEL-1-Fc promotes bone regeneration in 12-month-old mice (A) Experimental design. (B, left) Measurement of bone heights (distance from cement-enamel junction [CEJ] to alveolar bone crest [ABC]; CEJ-ABC) in groups of 2-month-old and 12-month-old mice after ten days of ligature placement (10dL) or after 10 days of ligature placement followed by five days without ligatures to enable resolution from periodontitis (10dL + 5dR). (B, middle) Data from the left panel were transformed to show bone loss at ligated (L) sites vs. unligated (U) contralateral sites. (B, right) Data from the middle panel were transformed to show bone growth relative to corresponding 10dL group (baseline). (C) Experimental design. (D) 12-month-old mice were subjected to LIP for ten days with or without ligature removal for five days to enable resolution, with or without local microinjection with DEL-1-Fc (1 μg) or equal molar amount of Fc control. Treatments were performed daily on days 10–14. (D, left) Measurement of bone heights (CEJ-ABC distance). (D, middle) Data from the left panel were transformed to show bone loss in ligated (L) sites vs. unligated (U) contralateral sites. (D, right) Data from the middle panel were transformed to indicate bone growth relative to the 10dL group (baseline). Data are means ± S.D. (B, n = 5–6 mice/group; D, n = 4–7 mice/group). ∗p < 0.05, ∗∗p < 0.01; one-way ANOVA and Dunnett’s test.

Article Snippet: Rabbit monoclonal anti-DEL-1 (clone EPR12451) , Abcam , Cat# ab190692.

Techniques: Transformation Assay

Journal: iScience

Article Title: A novel macrolide–Del-1 axis to regenerate bone in old age

doi: 10.1016/j.isci.2024.108798

Figure Lengend Snippet: Macrolides induce bone regeneration in a DEL-1-dependent manner in 18-month-old mice with naturally occurring periodontitis (A and B) Aged WT and aged Del1 −/− mice were administered macrolide antibiotics or vehicle control (ethanol) intraperitoneally daily for nine days. (C) Aged WT were administered macrolide antibiotics intraperitoneally twice a week for eight weeks. (A–C) The distance from the CEJ-ABC was measured (left panels). The CEJ-ABC data were transformed to show the bone changes relative to the control, which was set as the baseline (right panels). Positive values (in mm) indicate bone growth relative to the baseline (control). (D and E) Analysis of gene expression from gingiva samples collected from (D) aged WT and (E) aged Del1 −/− mice after nine days of drug treatment. The mRNA levels of the indicated genes were quantified by qPCR. Data were normalized to Gapdh mRNA and plotted relative to the aged control group, set as 1. (F and G) Coronal maxillary sections from (F) aged WT and (G) aged Del1 −/− mice were stained with modified Masson’s trichrome stain, which stains the mature (old) bone blue and new bone formation red . T: tooth; PDL: periodontal ligament; OB: old bone as blue; NB : new bone as red . Scale bars, 100 μm. Data are means ± SD (A, n = 8 mice/group; B, n = 4–6 mice/group; C, n = 6 mice/group; D-E, n = 5 mice/group). ∗∗p < 0.01; one-way ANOVA and Dunnett’s test.

Article Snippet: Rabbit monoclonal anti-DEL-1 (clone EPR12451) , Abcam , Cat# ab190692.

Techniques: Transformation Assay, Expressing, Staining, Modification

Journal: iScience

Article Title: A novel macrolide–Del-1 axis to regenerate bone in old age

doi: 10.1016/j.isci.2024.108798

Figure Lengend Snippet: Macrolides induce DEL-1 protein expression in PDL spaces of the aged periodontium and promote the binding of C/EBPβ to the DEL1 promoter Aged WT and aged Del1 −/− mice were administered macrolide antibiotics or vehicle control (ethanol) intraperitoneally daily for nine days. (A) Representative IF images of maxillary sections from aged WT mice obtained with fluorescence microscopy. Frozen maxillae sections were stained for DEL-1 (green), CD31 (red), and the nuclei were counterstained using DAPI (blue). T: Tooth; PDL: periodontal ligament; B: alveolar bone. Scale bars, 100 μm. (B) Mean fluorescence intensity (MFI) values were measured from randomly selected IF images from aged WT mice using the ImageJ software. All MFI values were normalized and plotted relative to the MFI of the control group, set as 1. (C) Representative IF images of maxillary sections from aged Del1 −/− mice obtained with fluorescence microscopy. Frozen maxillae sections were stained for DEL-1 (green), CD31 (red), and the nuclei were counterstained using DAPI (blue). T: tooth; PDL: periodontal ligament; B: alveolar bone. Scale bars, 100 μm. (D) The mRNA levels of DEL1 from hPDLCs were quantified by qPCR on day seven of the drug administration. Data were normalized to GAPDH mRNA and plotted relative to the aged control group, set as 1. (E) hPDLCs were transiently transfected with hEDIL3-promoter-Luc reporter plasmid, pre-treated for 30 min with ethanol control, or indicated macrolides (10 μg/mL), and analyzed for luciferase activity. A renilla luciferase construct was co-transfected as an internal control for normalization. Data are calculated as fold change relative to ethanol control treatment, which was set as 1. (F) ChIP analysis of C/EBPβ binding capacity at the DEL1 promoter in hPDLCs treated for 4 h with ethanol control or indicated macrolides (10 μg/mL). Non-immunoprecipitated cell extracts were used as input samples. Data are plotted as a percentage of input. Data are means ± SD (B, n = 6 mice/group; D, n = 3 sets of cultures/group; E, n = 6 sets of cultures/group; F, n = 4 sets of cultures/group). ∗p < 0.05, ∗∗p < 0.01; one-way ANOVA and Dunnett’s test.

Article Snippet: Rabbit monoclonal anti-DEL-1 (clone EPR12451) , Abcam , Cat# ab190692.

Techniques: Expressing, Binding Assay, Fluorescence, Microscopy, Staining, Software, Transfection, Plasmid Preparation, Luciferase, Activity Assay, Construct, Immunoprecipitation

Journal: iScience

Article Title: A novel macrolide–Del-1 axis to regenerate bone in old age

doi: 10.1016/j.isci.2024.108798

Figure Lengend Snippet: EM-523, a non-antibiotic erythromycin derivative, promotes bone regeneration in aged mice (A and B) Aged WT mice were treated with ethanol control, ERM, or EM-523 intraperitoneally (A) daily for nine days or (B) twice a week for eight weeks. The distance from the CEJ-ABC was measured (left panels). The CEJ-ABC data were transformed to show the bone changes relative to the control, which was set as the baseline (right panels). Positive values (in mm) indicate bone growth relative to the baseline (control). (C) Representative IF images of maxillary sections obtained by fluorescence microscopy. Frozen maxillae sections were stained for DEL-1 (green), CD31 (red), and the nuclei were counterstained using DAPI (blue). T: tooth; PDL: periodontal ligament; B: alveolar bone. Scale bars, 100 μm. (D) Mean fluorescence intensity (MFI) values of DEL-1 were measured from randomly selected IF images from aged WT using the ImageJ software. All MFI values were normalized and presented relative to the MFI of the control group in aged WT mice, set as 1. (E) The mRNA levels of DEL1 from human periodontal ligament cells (hPDLCs) were quantified by qPCR on day seven after drug treatment. Data were normalized to GAPDH mRNA and plotted relative to the aged control group, set as 1. (F) hPDLCs were transiently transfected with hDEL1-promoter-Luc reporter plasmid, pre-treated for 30 min with ethanol control, ERM (10 μg/mL), or EM-523 (1 μg/mL), and analyzed for luciferase activity. A renilla luciferase construct was co-transfected as an internal control for normalization. Data were calculated as fold change relative to ethanol control treatment, which was set as 1. (G) ChIP analysis of C/EBPβ binding capacity at the DEL1 promoter in hPDLCs treated for 4 h with ethanol control, ERM, or EM-523 (10 and 1 μg/mL, respectively). Non-immunoprecipitated cell extracts were used as input samples. Data are plotted as a percentage of input. (H) Gene expression analysis using qPCR of primary mouse PDLCs on day nine of the osteogenic differentiation assay for the expression of the indicated genes. Cells were treated with vehicle control, macrolides, or EM-523 (ERM, CLR and AZM as 10 μg/mL; EM-523 as 1 μg/mL). Data were normalized to Gapdh mRNA and plotted relative to ethanol-treated control, set as 1. (I–N) Aged WT mice were treated with ethanol control, ERM, or EM-523 intraperitoneally for nine days. (I) Optical microscopy images of WT maxillary sections show PDL areas stained for ALP activity. T: Tooth; PDL: periodontal ligament; B: alveolar bone. Scale bars, 100 μm. (J) The ALP-positive region of each area was quantified and represented as a percentage of the total area. (K) Optical microscopy images of WT maxillary sections show PDL areas stained for TRAP. Arrowheads indicate TRAP+ cells in PDL space. T: Tooth; PDL: periodontal ligament; B: alveolar bone. Scale bars, 100 μm. (L) The number of TRAP + cells within the PDL space were counted and compared between indicated groups. (M) Representative IF images of maxillary sections obtained by fluorescence microscopy. Frozen maxillary sections were stained for alpha-smooth muscle actin (α-SMA, green), and the nuclei were counterstained using DAPI (blue). T: tooth; PDL: periodontal ligament; B: alveolar bone. Scale bars, 100 μm. (N) Mean fluorescence intensity (MFI) values were measured from IF images using the ImageJ software. All MFI values were normalized and plotted relative to the MFI of the control group in aged mice, set as 1. (O) Cultures of hPDLCs were treated with ethanol control, ERM, or EM-523 (10 and 1 μg/mL). The mRNA levels of ACTA2 from hPDLCs were quantified by qPCR on day seven after drug treatment. Data were normalized to GAPDH mRNA and plotted relative to the aged control group, set as 1. Data are means ± S.D. (A, n = 8–10 mice/group; B, n = 6 mice/group; D, n = 6 mice/group; E, n = 3 sets of cultures/group; F, n = 6 sets of cultures/group; G, n = 4 sets of cultures/group; H, n = 3–4 sets of cultures/group; J, n = 6 mice/group; L, n = 9 mice/group; N, n = 6 mice/group; O, n = 5 sets of cultures/group). ∗p < 0.05, ∗∗p < 0.01; (one-way ANOVA and Tukey’s test; only H, one-way ANOVA and Bonferroni’s test).

Article Snippet: Rabbit monoclonal anti-DEL-1 (clone EPR12451) , Abcam , Cat# ab190692.

Techniques: Transformation Assay, Fluorescence, Microscopy, Staining, Software, Transfection, Plasmid Preparation, Luciferase, Activity Assay, Construct, Binding Assay, Immunoprecipitation, Expressing, Differentiation Assay

Journal: iScience

Article Title: A novel macrolide–Del-1 axis to regenerate bone in old age

doi: 10.1016/j.isci.2024.108798

Figure Lengend Snippet: EM-523 promotes bone regeneration and suppresses osteoclastogenesis in vitro (A) MC3T3-E1 cells were cultured in osteogenic differentiation medium treated with control solvent or indicated macrolides. (A, left panel) Representative images of mineralization nodules from osteoblasts, stained with Alizarin Red S (ARS) staining after 26 days of the osteogenic differentiation assay. (A, middle panel) The mineralization area in each culture was quantified and represented as a percentage of the total area. (A, right panel) Quantification of the ARS staining using 10% cetylpyridinium chloride (CPC) on day 26 of the osteogenic differentiation assay. The absorbance was measured at 560 nm. The control group was incubated with an osteogenic differentiation medium only. (B) MC3T3-E1 cells were cultured in osteogenic differentiation medium treated with ethanol, ERM, or EM-523. (B, left panel) Representative images of mineralization nodules from osteoblasts, stained with ARS after 20 days of the osteogenic differentiation assay. (B, right panel) The mineralization area in each culture was quantified and presented as a percentage of the total area. The control group was incubated with an osteogenic differentiation medium only. (C) hPDLCs were cultured in osteogenic differentiation medium with control solvent or indicated macrolides. (C, left panel) Representative images of hPDLCs cultures, stained with alkaline phosphatase stain after 26 days of the osteogenic differentiation assay. Scale bars, 200 μm. (C, right panel) The ALP-positive region of each culture was quantified and represented as a percentage of the total area. (D) Gene expression analysis of hPDLCs on day nine of the osteogenic differentiation assay using qPCR. RUNX2 , SP7 , and BGLAP , as representative markers for early, middle, and late osteogenic markers, respectively. Data were normalized to GAPDH mRNA and plotted relative to ethanol-treated control, set as 1. (E) hMSCs were cultured in osteogenic differentiation medium with ethanol or indicated macrolides (10 μg/mL). (E, upper panel) Representative images of mineralization nodules from osteoblasts, stained with Alizarin Red S after 26 days of the osteogenic differentiation assay. (E, middle left panel) The mineralization area in each culture was quantified and presented as a percentage of the total area. (E, middle right panel) Quantification of the Alizarin Red S staining using 10% CPC on day 26 of the osteogenic differentiation assay. (E, lower panel) Gene expression analysis using qPCR of human MSCs on day nine of the osteogenic differentiation assay for the expression of the indicated genes. Data were normalized to GAPDH mRNA and plotted relative to ethanol control, set as 1. (F) Bone marrow cells were collected from WT mice for the osteoclastogenesis assay. Cells were treated with DEL-1-Fc (1 μg/mL), equal molar amount of Fc control, control solvent, or indicated macrolides for one week. (F, upper panel) Representative images of the culture well from each group after TRAP staining are shown. Histological images of TRAP + MNCs. Scale bars, 200 μm. (F, lower left panel) The average size of TRAP + MNCs was measured using the ImageJ software. (F, lower right panel) The percentage of TRAP + area per total area was measured using the ImageJ software. (G) Primary osteoblastic progenitor cells were collected from Del1 −/− mice. Primary osteoblastic progenitor cells were cultured in osteogenic differentiation media administered with DEL-1-Fc, equal molar amount of Fc control, control solvent, or indicated macrolides for 26 days. (G, upper panel) Representative images of mineralization nodules from osteoblasts, stained with Alizarin Red S after 26 days of the osteogenic differentiation assay. (G, lower panel) Gene expression analysis was completed on day nine of the osteogenic differentiation assay using qPCR. Runx2 , Sp7 , and Bglap , as representative markers for early, middle, and late osteogenic markers, respectively. Data were normalized to GAPDH mRNA and plotted relative to ethanol control, set as 1. (H) Bone marrow cells were collected from Del1 −/− mice for the osteoclastogenesis assay. Cells were treated with DEL-1-Fc, equal molar amount of Fc control, control solvent, or indicated macrolides for one week. (H, upper panel) Representative images of the culture well from each group after TRAP staining were shown. Histological images of TRAP + MNCs. Scale bars, 200 μm. (H, lower left panel) The average size of TRAP + MNCs was measured using the ImageJ software. (H, lower right panel) The percentage of TRAP + area per total area was measured using the ImageJ software. Data are means ± S.D. (A–C, n = 4 sets of cultures/group; D, n = 5 sets of cultures/group; E middle and lower panel, n = 4 sets of cultures/group; F–H, n = 5 sets of cultures/group). ∗p < 0.05, ∗∗p < 0.01; one-way ANOVA and Bonferroni’s test.

Article Snippet: Rabbit monoclonal anti-DEL-1 (clone EPR12451) , Abcam , Cat# ab190692.

Techniques: In Vitro, Cell Culture, Solvent, Staining, Differentiation Assay, Incubation, Expressing, Software

Journal: iScience

Article Title: A novel macrolide–Del-1 axis to regenerate bone in old age

doi: 10.1016/j.isci.2024.108798

Figure Lengend Snippet:

Article Snippet: Rabbit monoclonal anti-DEL-1 (clone EPR12451) , Abcam , Cat# ab190692.

Techniques: Recombinant, Construct, Transfection, Staining, Chromatin Immunoprecipitation, Magnetic Beads, Luciferase, SYBR Green Assay, Generated, Binding Assay, Expressing, Plasmid Preparation, Software, Imaging, Microscopy, Fluorescence, Real-time Polymerase Chain Reaction