Journal: Bone Research

Article Title: Bone marrow adipogenic lineage precursors are the major regulator of bone resorption in adult mice

doi: 10.1038/s41413-025-00405-4

Figure Lengend Snippet: Depletion of RANKL in MALPs increases long bone trabecular bone mass in adult mice by suppressing bone resorption. a qRT-PCR analysis of Rankl mRNA in bone marrow and cortical bone from WT and RANKL iCKO mice at 4 weeks after Tam injection. Mice received Tam at 3 months of age. n = 3 mice/group. b ELISA analysis of RANKL in bone marrow from WT and RANKL iCKO mice at 2 weeks after Tam injection. Mice received Tam at 3 months of age. n = 3 mice/group. c 3D microCT reconstruction of whole femurs from WT and iCKO mice at 1 month after Tam injection. Scale bar = 1 mm. d 3D microCT reconstruction reveals a drastic increase of femoral trabecular bone. Scale bar = 200 µm. e MicroCT measurement of trabecular bone structural parameters. BV/TV bone volume fraction, Tb.N trabecular number, Tb.Th trabecular thickness, Tb.Sp trabecular separation. f Representative TRAP staining images show TRAP+ osteoclast (arrows) at different skeletal sites: secondary spongiosa (SS), chondro-osseous junction (COJ), and endosteal surface (Endo.S). Scale bar = 50 μm. g Quantification of osteoclast surface (Oc.S) at 3 skeletal sites. BS bone surface, L COJ length. h Representative Osterix staining of trabecular bone from WT and RANKL iCKO femurs. Scale bar = 50 μm. i Quantification of osteoblast surface (OB.S). j Representative double labeling of trabecular bone from WT and iCKO femurs. Scale bar = 20 μm. k Bone formation activity is quantified. MAR mineral apposition rate, MS mineralizing surface, BFR bone formation rate. l Serum ELISA analysis of bone resorption marker (CTX-1) and formation marker (P1NP) in WT and iCKO mice. * P < 0.05; ** P < 0.01; *** P < 0.001 vs WT , n = 5–6 mice/group

Article Snippet: Sera were collected during mouse euthanization for measuring bone turnover markers, collagen type I C-telopeptide degradation products (mouse CTX-I ELISA Kit, MyBioSource), N-terminal propeptide of type I procollagen (ImmunotagTM Mouse PINP ELISA Kit, G-Bioscience) according to the manufacturer’s instructions.

Techniques: Quantitative RT-PCR, Injection, Enzyme-linked Immunosorbent Assay, Staining, Labeling, Activity Assay, Marker

Journal: Bone Research

Article Title: Bone marrow adipogenic lineage precursors are the major regulator of bone resorption in adult mice

doi: 10.1038/s41413-025-00405-4

Figure Lengend Snippet: RANKL deficiency in MALPs protects adult female mice from ovariectomy-induced trabecular bone loss. a 3D microCT reconstruction of femoral trabecular bone from WT and RANKL iCKO mice at 6 weeks post OVX surgery. Mice received Tam injections at 3 months of age right before the surgery. Scale bar = 200 µm. b MicroCT measurement of trabecular bone structural parameters. c Representative TRAP staining images of trabecular bone from WT and RANKL iCKO femurs show TRAP + osteoclasts (arrows). Scale bar = 50 μm. d Quantification of osteoclast surface (Oc.S). e Representative Osterix staining of trabecular bone from WT and RANKL iCKO femurs. Scale bar = 50 μm. f Quantification of osteoblast surface (OB.S). g Bone formation activity is quantified. h Serum ELISA analysis of bone resorption marker (CTX-1) and formation marker (P1NP) in WT and iCKO mice. i Representative H&E staining of trabecular bone from WT and RANKL iCKO femurs. Scale bar = 50 μm. j Quantification of the percentage of adipocyte area within bone marrow and adipocyte size. # P < 0.05; ## P < 0.01; ### P < 0.001 OVX vs Sham; * P < 0.05; ** P < 0.01; *** P < 0.001 iCKO vs WT , n = 5–6 mice/group

Article Snippet: Sera were collected during mouse euthanization for measuring bone turnover markers, collagen type I C-telopeptide degradation products (mouse CTX-I ELISA Kit, MyBioSource), N-terminal propeptide of type I procollagen (ImmunotagTM Mouse PINP ELISA Kit, G-Bioscience) according to the manufacturer’s instructions.

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

Journal: Bone Research

Article Title: Bone marrow adipogenic lineage precursors are the major regulator of bone resorption in adult mice

doi: 10.1038/s41413-025-00405-4

Figure Lengend Snippet: Depleting RANKL in MALPs in osteoporotic mice restores trabecular bone mass. a 3D microCT reconstruction of femoral trabecular bone from WT and RANKL iCKO mice at 10 weeks post OVX surgery. Mice received the surgery at 3 months of age and vehicle or Tam injections 6 weeks later. Scale bar = 200 µm. b MicroCT measurement of trabecular bone structural parameters. c Representative TRAP staining images of femoral trabecular bone from WT and RANKL iCKO mice with vehicle or Tam injections show TRAP + osteoclasts (arrows). Scale bar = 50 μm. d Quantification of osteoclast surface (Oc.S). e Representative Osterix staining of femoral trabecular bone from WT and RANKL iCKO mice with vehicle or Tam injections. Scale bar = 50 μm. f Quantification of osteoblast surface (OB.S). g Bone formation activity is quantified. h Serum ELISA analysis of bone resorption marker (CTX-1) and formation marker (P1NP) in WT and iCKO mice with vehicle or Tam injections. # P < 0.05; ## P < 0.01; ## P < 0.001 Tam vs Veh; * P < 0.05; ** P < 0.01; *** P < 0.001 iCKO vs WT , n = 5–6 mice/group

Article Snippet: Sera were collected during mouse euthanization for measuring bone turnover markers, collagen type I C-telopeptide degradation products (mouse CTX-I ELISA Kit, MyBioSource), N-terminal propeptide of type I procollagen (ImmunotagTM Mouse PINP ELISA Kit, G-Bioscience) according to the manufacturer’s instructions.

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