Journal: eLife

Article Title: Integrin alpha11 is an Osteolectin receptor and is required for the maintenance of adult skeletal bone mass

doi: 10.7554/eLife.42274

Figure Lengend Snippet: ( A ) Lepr-Cre; Itga11 fl/fl mice were grossly normal and indistinguishable from littermate controls. ( B – D ) Body length ( B ), body mass ( C ) and femur length ( D ) did not significantly differ between Lepr-Cre; Itga11 fl/fl mice and sex-matched littermate controls at 2, 6, or 12 months of age (n = 4–7 mice per genotype per sex per time point, from at least three independent experiments). ( E ) ELISA measurement of serum Osteolectin levels in Lepr-Cre; Itga11 fl/fl mice and littermate controls at 2, 6, and 12 months of age (n = 8 mice per genotype per time point from four independent experiments). ( F ) Representative microCT images of trabecular bone in the distal femur metaphysis of male Lepr-Cre; Itga11 fl/fl mice and littermate controls at 2, 6, and 12 months of age. ( G–L ) microCT analysis of trabecular bone volume/total volume ( G ), trabecular number ( H ), trabecular bone thickness ( I ), trabecular bone spacing ( J ), connectivity density ( K ), and bone mineral density ( L ) in the distal femur metaphysis of Lepr-Cre; Itga11 fl/fl mice and sex-matched littermate controls at 2, 6, and 12 months of age (n = 4–7 mice per genotype per sex per time point from at least three independent experiments). ( M ) Trabecular bone mineral apposition rate based on calcein double labelling in the distal femur metaphysis (n = 3 mice per genotype per sex per time point). ( N ) Bone resorption rate analysis by measuring the deoxypyridinoline/creatinine ratio in the urine of LepR-Cre; Itga11 fl/fl mice and littermate controls at 6 and 12 months of age (n = 4–5 mice per genotype per time point from three independent experiments). ( O ) Serum P1NP levels in LepR-Cre; Itga11 fl/fl mice and sex-matched littermate controls at 6 and 12 months of age (n = 4 mice per genotype per time point from three independent experiments). All numerical data reflect mean ±standard deviation. The statistical significance of differences was determined with two-way ANOVAs with Sidak’s multiple comparisons tests. 10.7554/eLife.42274.012 Figure 5—source data 1. Data for Data for .

Article Snippet: Commercial assay or kit , Rat/Mouse PINP ELISA kit , Immunodiagnostic Systems , AC-33F1 , .

Techniques: Enzyme-linked Immunosorbent Assay, Standard Deviation

Journal: eLife

Article Title: Integrin alpha11 is an Osteolectin receptor and is required for the maintenance of adult skeletal bone mass

doi: 10.7554/eLife.42274

Figure Lengend Snippet:

Article Snippet: Commercial assay or kit , Rat/Mouse PINP ELISA kit , Immunodiagnostic Systems , AC-33F1 , .

Techniques: Recombinant, Diagnostic Assay, Cell Culture, Protease Inhibitor, Western Blot, Reverse Transcription, Enzyme-linked Immunosorbent Assay, Fractionation

Journal: The Journal of Clinical Investigation

Article Title: Bone marrow adipogenic lineage precursors promote osteoclastogenesis in bone remodeling and pathologic bone loss

doi: 10.1172/JCI140214

Figure Lengend Snippet: (A) Representative TRAP staining images show TRAP+ osteoclasts (arrowheads) at different skeletal sites: secondary spongiosa (ss), COJ, and endosteal surface (Endo.S). Scale bar: 50 μm. (B) Quantification of osteoclast surface (Oc.S) and osteoclast number (Oc.N) at 3 skeletal sites (n = 5–6 mice/group). BS, bone surface; L, COJ length. (C) Quantification of osteoblast number (Ob.N) in the secondary spongiosa and at the endosteal surface (n = 5–6 mice/group). (D) Quantification of osteocyte density (osteocyte number per bone area, Ocy.N/BA) in the secondary spongiosa (n = 5–6 mice/group). (E) Representative double labeling in distal femurs of WT and CKO mice. Scale bar: 10 μm. (F) Bone formation activity is quantified (n = 5–6/group). (G) Serum ELISA analysis of bone resorption marker (CTX-1) and formation marker (PINP) in WT and CKO mice (n = 5 mice/group). *P < 0.05; **P < 0.01; ***P < 0.001 CKO vs. WT, 2-tailed unpaired Student’s t test.

Article Snippet: Sera were collected from WT and RANKL-CKO Adipoq mice during euthanization for measuring bone turnover markers, collagen type I C-telopeptide degradation products (CTX-I RatLaps EIA; Immunodiagnostic Systems) and N-terminal propeptide of type I procollagen (PINP ELISA Kit, MyBioSource) respectively, according to the manufacturer’s instructions.

Techniques: Staining, Labeling, Activity Assay, Enzyme-linked Immunosorbent Assay, Marker

Journal: iScience

Article Title: The inhibition of PINK1/Drp1-mediated mitophagy by hyperglycemia leads to impaired osteoblastogenesis in diabetes

doi: 10.1016/j.isci.2024.111519

Figure Lengend Snippet: BMP9 promoted mitophagy markers to the control levels in diabetic mice in vivo (A) Representative images derived from micro-CT analysis, including 2D image construction of distal femur, 3D images reconstruction of trabecular bone of distal femur, and 3D image reconstruction of the femoral midshaft corticoid bone. (B–D) Quantitative analysis of the vBMD, BV/TV and Ct.Th of corticoid bone by micro-CT. (E–I) Quantitative analysis of the BV/TV, Tb.N, Tb.Sp, Tb.Th, and vBMD of trabecular by micro-CT. (J) Representative images of HE-stained decalcified femur sections. Scale bars, 200 μm. (K–M) The right femur was isolated and subjected to biomechanical properties analysis. The maximum load, elastic modulus and SMI were evaluated for each group. (N) Representative images derived from micro-CT analysis, including 2D image construction and 3D image reconstruction of L3 lumbar vertebra. (O–S) Quantitative analysis of the vBMD, BV/TV, Tb.N, Tb.Th, and Tb.Sp of L3 by micro-CT. (T and U) The levels of serum bone turnover parameters PINP and CTX-I were detected by ELISA. (V) WB analysis of protein levels of mitophagy marker in skull. (W) Expressions of ALP, Osx, Drp1, and PINK1 in skull were determined by RT-qPCR. (X) Immunofluorescence analysis of Drp1 and PINK1 expression in femur sections. Scale bars, 10 μm. (Y) Immunofluorescence analysis of PINK1 expression in femur sections. Scale bars, 20 μm. (Z) Immunofluorescence results of double labeling of osteoblasts with PINK1 and Drp1 in femur sections. Scale bars, 20 μm. Data presented as mean ± SD. n = 8 biological replicates. One-way ANOVA was used for comparison among multiple groups. ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001. Ns, no significance.

Article Snippet: ELISA Kit for Procollagen I N-Terminal Propeptide (PINP) , USCN Life Science , Cat# SEA957Hu.

Techniques: Control, In Vivo, Derivative Assay, Micro-CT, Staining, Isolation, Enzyme-linked Immunosorbent Assay, Marker, Quantitative RT-PCR, Immunofluorescence, Expressing, Labeling, Comparison

Journal: iScience

Article Title: The inhibition of PINK1/Drp1-mediated mitophagy by hyperglycemia leads to impaired osteoblastogenesis in diabetes

doi: 10.1016/j.isci.2024.111519

Figure Lengend Snippet:

Article Snippet: ELISA Kit for Procollagen I N-Terminal Propeptide (PINP) , USCN Life Science , Cat# SEA957Hu.

Techniques: Recombinant, Staining, Isolation, Bicinchoninic Acid Protein Assay, ROS Assay, Enzyme-linked Immunosorbent Assay, RNA Sequencing, Sequencing, Expressing, Plasmid Preparation, Software, Real-time Polymerase Chain Reaction, Spectrophotometry, Microscopy

Journal: Bone Research

Article Title: HuR-mediated nucleocytoplasmic translocation of HOTAIR relieves its inhibition of osteogenic differentiation and promotes bone formation

doi: 10.1038/s41413-023-00289-2

Figure Lengend Snippet: HOTAIR overexpression in MSCs delays bone development. a , b Alcian blue and Alizarin red staining of the whole skeletons of WT or Prx1-HOTAIR TG mice at 5 days old. Scale bar, 1 cm. c Representative images showing three-dimensional trabecular architecture by micro-CT reconstruction from WT and Prx1-HOTAIR TG male mice at 1 month old. Scale bar, 0.5 mm. d Representative images showing the three-dimensional trabecular architecture and cortical architecture as shown by micro-CT reconstruction at the distal femurs from WT and Prx1-HOTAIR TG mice at 1 month old. Scale bar, 0.5 mm. e Micro-CT measurements of BV/TV, BMD, Tb.N, Tb.Th, Tb.Sp, and Cort.Th in the distal femurs of mice. n = 10 for each group. BV/TV, ratio of bone volume to tissue volume; BMD, bone mineral density; Tb.N, trabecular number; Tb.Th, trabecular thickness; Tb.Sp, trabecular separation; Cort.Th, cortical bone thickness. f Representative images showing new bone formation assessed by double calcein labeling in WT and Prx1-HOTAIR TG mice. Scale bar, 20 μm. g Quantification of mineral apposition rate (MAR) and osteoblast number to bone surface (N.ob/BS) ( n = 6 for each group). h The maximal (max.) load at failure determined by three-point bending of femurs from WT ( n = 10) and Prx1-HOTAIR TG ( n = 10) mice. i Histological images for Ocn staining of the proximal tibia from WT and Prx1-HOTAIR TG mice. Scale bar, 100 μm. j ELISA analysis of the PINP protein level in the serum from WT ( n = 10) and Prx1-HOTAIR TG ( n = 10) mice. k qRT-PCR analysis of Alp , Runx2 and Sp 7 mRNA levels in bone tissues collected from WT ( n = 10) and Prx1-HOTAIR TG ( n = 10) mice. Two-tailed unpaired Student’s t test was used for statistical evaluations of two group comparisons. All data are the mean ± s.e.m. *P < 0.05, **P < 0.01, ***P < 0.001

Article Snippet: The serum levels of PINP were detected by a mouse PINP (procollagen type I N-terminal propeptide) ELISA kit (ImmunoWay, KE1744) according to the product instructions.

Techniques: Over Expression, Staining, Micro-CT, Labeling, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR, Two Tailed Test

Journal: Bone Research

Article Title: HuR-mediated nucleocytoplasmic translocation of HOTAIR relieves its inhibition of osteogenic differentiation and promotes bone formation

doi: 10.1038/s41413-023-00289-2

Figure Lengend Snippet: HOTAIR overexpression in osteoblasts increases bone formation. a Schematic representation of the transgenic construct used to generate osteoblast-specific HOTAIR overexpression transgenic mouse lines. b qRT‒PCR analysis of HOTAIR levels in bone and other tissues from 6-month-old WT and osteoblast-specific HOTAIR-overexpressing mice (Bglap-HOTAIR TG) ( n = 6). c Representative images showing three-dimensional trabecular architecture by micro-CT reconstruction at 6 months of age. Scale bar, 0.5 mm. d Representative images showing the three-dimensional trabecular architecture and cortical architecture as shown by micro-CT reconstruction at the distal femurs at 6 months of age. Scale bar, 0.5 mm. e Micro-CT measurements of BV/TV, BMD, Tb.N, Tb.Th, Tb.Sp, and Cort.Th in the distal femurs of mice ( n = 10 for each group). f Representative images showing new bone formation assessed by double calcein labeling. Scale bar, 20 μm. g Quantification of mineral apposition rate (MAR) and osteoblast number to bone surface (N.ob/BS) ( n = 6 for each group). h The maximal (max.) load at failure determined by three-point bending of femurs from WT ( n = 10) and Ocn-HOTAIR TG ( n = 10) mice. i Histological images for Ocn staining of the proximal tibia. Scale bar, 100 μm. j ELISA analysis of the PINP protein level in the serum from WT ( n = 10) and Bglap-HOTAIR TG ( n = 10) mice. k qRT-PCR analysis of Alp , Bglap and Col1a1 mRNA levels in bone tissues collected from WT ( n = 10) and Bglap-HOTAIR TG ( n = 10) mice. Two-tailed unpaired Student’s t test was used for statistical evaluations of two group comparisons. All data are the mean ± s.e.m. *P < 0.05, **P < 0.01, ***P < 0.001

Article Snippet: The serum levels of PINP were detected by a mouse PINP (procollagen type I N-terminal propeptide) ELISA kit (ImmunoWay, KE1744) according to the product instructions.

Techniques: Over Expression, Transgenic Assay, Construct, Micro-CT, Labeling, Staining, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR, Two Tailed Test

Journal: Bone Research

Article Title: HuR-mediated nucleocytoplasmic translocation of HOTAIR relieves its inhibition of osteogenic differentiation and promotes bone formation

doi: 10.1038/s41413-023-00289-2

Figure Lengend Snippet: Osteoblast-specific overexpression of HOTAIR alleviates unloading-induced bone loss. a Representative image showing three-dimensional distal femur trabecular architecture by micro-CT reconstruction from the indicated groups of mice. Representative images of six independent tissues in each group. Ctrl, control group, HU, hindlimb unloading group. Scale bar, 0.5 mm. b Micro-CT measurements for BV/TV, BMD, Tb.N and Tb.Sp at the distal femurs. n = 10 for each group. c Representative images of Ocn staining of the proximal tibia. Scale bar, 100 μm. d ELISA analysis of PINP protein levels in the serum. n = 10 for each group. e qRT-PCR analysis of Alp , Bglap , and Col1a1 mRNA levels in bone tissues. n = 10 for each group. f , g qRT-PCR analysis of HOTAIR and miR-214 levels in bone tissues. n = 10 for each group. h Western blot analysis of Atf4 protein levels in bone tissue. All data are the mean ± s.e.m. Statistical analysis with more than two groups was performed with two-way analysis of variance (ANOVA) with the Šídák post hoc test to determine group differences. All data are the mean ± s.e.m. *P < 0.05, **P < 0.01, ***P < 0.001

Article Snippet: The serum levels of PINP were detected by a mouse PINP (procollagen type I N-terminal propeptide) ELISA kit (ImmunoWay, KE1744) according to the product instructions.

Techniques: Over Expression, Micro-CT, Control, Staining, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR, Western Blot

Journal: eLife

Article Title: The mechanosensitive Piezo1 channel is required for bone formation

doi: 10.7554/eLife.47454

Figure Lengend Snippet: ( a ) Alcian blue and Alizarin red staining of the skeletons of Piezo1 fl/fl or Piezo1 Ocn/Ocn mice at 7 days old. ( b, c ) Representative images showing the three-dimensional cortical bone and trabecular architecture as shown by micro-CT reconstruction at the distal femurs from Piezo1 fl/fl or Piezo1 Ocn/Ocn mice at 2 months old. Scale bars: ( b ) 1.0 mm, ( c ) 0.5 mm. ( d ) Micro-CT measurements for bone mineral density (BMD), trabecular bone volume fraction (BV/TV), trabecular number (Tb.N), trabecular thickness (Tb.Th), trabecular separation (Tb.Sp) and cortical thickness (Cort.Th) at the distal femurs from Piezo1 fl/fl (n = 6) or Piezo1 Ocn/Ocn (n = 7) mice. ( e ) Relative maximal (max.) load at failure determined by three-point bending of femurs from Piezo1 fl/fl (n = 6) and Piezo1 Ocn/Ocn mice (n = 7). ( f ) Representative images showing new bone formation assessed by double calcein labeling in Piezo1 fl/fl mice (n = 6) and Piezo1 Ocn/Ocn mice (n = 7). MAR, mineral apposition rate. BFR/BS, bone formation rate/bone surface. Scale bar, 100 μm. ( g ) Histology images for Col1α1 and Ocn staining of the proximal tibia from Piezo1 fl/fl and Piezo1 Ocn/Ocn mice. Scale bar, 25 μm. ( h ) ELISA analysis of the levels of PINP and Ocn protein levels in the serum from Piezo1 fl/fl (n = 8) and Piezo1 Ocn/Ocn mice (n = 9). ( i ) QRT-PCR analysis of Alp , Bglap and Col1α1 mRNA levels in bone tissues collected from Piezo1 fl/fl (n = 6) and Piezo1 Ocn/Ocn mice (n = 7). All data are the mean ± s.e.m. *, p<0.05; **, p<0.01. 10.7554/eLife.47454.013 Figure 3—source data 1. The data and statistical analysis of Piezo1 Ocn/Ocn mice show severely impaired bone formation.

Article Snippet: Commercial assay or kit , Mouse PINP ELISA Kit , Immunoway , KE1744 , .

Techniques: Staining, Micro-CT, Labeling, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR

Journal: eLife

Article Title: The mechanosensitive Piezo1 channel is required for bone formation

doi: 10.7554/eLife.47454

Figure Lengend Snippet: ( a ) The appearance and body length of 2-month-old Piezo1 fl/fl and Piezo1 Ocn/Ocn mice. ( b ) The body weight of Piezo1 fl/fl (n = 11) and Piezo1 Ocn/Ocn mice (n = 11). ( c ) The appearance and the length of femurs and tibias from Piezo1 fl/fl (n = 7) and Piezo1 Ocn/Ocn mice (n = 7). ( d ) Representative images of Trap staining in bone tissue from Piezo1 fl/fl and Piezo1 Ocn/Ocn mice. N.Oc/B.Pm, osteoclast number/bone perimeter. ( e ) ELISA analysis of the levels of CTX-1 protein level in the serum from Piezo1 fl/fl (n = 10) and Piezo1 Ocn/Ocn mice (n = 9). ( f ) QRT-PCR analysis of Nfatc1, Acp5, Ctsk and Mmp9 mRNA levels in bone tissue from Piezo1 fl/fl (n = 6) and Piezo1 Ocn/Ocn mice (n = 8). All data are the mean ± s.e.m. **, p<0.01.

Article Snippet: Commercial assay or kit , Mouse PINP ELISA Kit , Immunoway , KE1744 , .

Techniques: Staining, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR

Journal: eLife

Article Title: The mechanosensitive Piezo1 channel is required for bone formation

doi: 10.7554/eLife.47454

Figure Lengend Snippet: ( a ) Representative images showing three-dimensional trabecular architecture as determined by micro-CT reconstruction of the distal femurs from the groups of mice indicated. Scale bar, 0.5 mm. ( b ) Micro-CT measurements for BMD, BV/TV, Tb.N and Tb.Th in the distal femurs from the groups of mice indicated. n = 6 in each group. ( c ) Relative maximal (max.) load at failure determined by three-point bending of femurs from the groups of mice indicated. Ctrl-Piezo1 fl/fl group, n = 9; HS-Piezo1 fl/fl group, n = 7; Ctrl- Piezo1 Ocn/Ocn group, n = 7; HS-Piezo1 Ocn/Ocn group, n = 8. ( d ) Histology images for Col1α1 and Ocn staining of the proximal tibia from the groups of mice indicated. Scale bar: 25 μm. ( e ) ELISA analysis of the levels of PINP and Ocn proteins in serum from the groups of mice indicated. Ctrl- Piezo1 fl/fl group, n = 9; HS-Piezo1 fl/fl group, n = 7; Ctrl-Piezo1 Ocn/Ocn group, n = 7; HS-Piezo1 Ocn/Ocn group, n = 10. ( f ) QRT-PCR analysis of Alp , Bglap and Col1α1 mRNA levels in bone tissues collected from the groups of mice indicated. n = 6 in each group. All data are the mean ± s.e.m. *, p<0.05; **, p<0.01; ***, p<0.001. 10.7554/eLife.47454.015 Figure 4—source data 1. The data and statistical analysis of the effect of mechanical unloading on bone remodeling and osteoblast function in Piezo1 fl/fl and Piezo1 Ocn/Ocn mice.

Article Snippet: Commercial assay or kit , Mouse PINP ELISA Kit , Immunoway , KE1744 , .

Techniques: Micro-CT, Staining, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR

Journal: eLife

Article Title: The mechanosensitive Piezo1 channel is required for bone formation

doi: 10.7554/eLife.47454

Figure Lengend Snippet:

Article Snippet: Commercial assay or kit , Mouse PINP ELISA Kit , Immunoway , KE1744 , .

Techniques: Recombinant, Plasmid Preparation, Sequencing, Enzyme-linked Immunosorbent Assay, Modification, Software