Journal: iScience

Article Title: CGK733 alleviates ovariectomy-induced bone loss through blocking RANKL-mediated Ca 2+ oscillations and NF-κB/MAPK signaling pathways

doi: 10.1016/j.isci.2023.107760

Figure Lengend Snippet:

Article Snippet: Mouse CTX-I ELISA Kit , Sangon Biotech , D721204.

Techniques: Recombinant, SYBR Green Assay, CCK-8 Assay, Reverse Transcription, ALP Assay, Enzyme-linked Immunosorbent Assay, Software

Journal: EMBO Reports

Article Title: Glutaminolysis provides nucleotides and amino acids to regulate osteoclast differentiation in mice

doi: 10.1038/s44319-024-00255-x

Figure Lengend Snippet: ( A ) Western blot analysis of LysMCre;Gls fl/+ (WT) and LysMCre;Gls fl/fl (cKO) BMM cultured with or without RANKL for 72 h ( n = 3). ( B – D ) TRAP staining ( B , C ) or qPCR ( D ) of primary BMM isolated from LysMCre;Gls fl/+ and LysM;Gls fl/fl female mice and cultured with RANKL for 4 days ( n = 5 independent experiments). Scale bar: 500 μm. ( C ) Quantification of the number of TRAP-positive multinucleated osteoclasts per well ( n = 5). ( D ) qPCR showing reduced induction of osteoclast genes in LysM;Gls fl/fl BMM induced with RANKL for 4 days ( n = 3 independent experiments). ( E ) Phalloidin staining showing reduced actin belt formation in LysM;Gls fl/fl BMM ( n = 4). Scale bar: 100 μm. ( F ) Quantification of the number of nuclei per osteoclast ( n = 5) and the number of actin belts ( n = 4). ( G – H ) Representative μCT ( G ) and calculated bone volume per tissue volume (BV/TV) ( H ) from the distal femur of 4-month-old LysMCre;Gls fl/+ and LysM;Gls fl/fl female mice ( n = 8 mice). Scale bar: 200 μm. ( I – J ) Representative TRAP staining ( I ) and quantification of osteoclast number per bone surface (Oc.N/BS) ( J ) from distal femurs from 4-month-old LysMCre;Gls fl/+ and LysM;Gls fl/fl female mice ( n = 5). Scale bar: 100 μm. ( K – L ) Serum ELISA for TRAP 5b ( K ) and CTX-I ( L ) from 4-month-old LysMCre;Gls fl/+ and LysM;Gls fl/fl female mice ( n = 8). ( M , N ) Representative H&E staining ( n = 7) and quantification of osteoblast number per bone surface (Ob.N/BS). Scale bar: 100 μm. ( O ) Serum ELISA for P1NP ( n = 8). Data are shown as mean ± SD. Two-tailed Student’s unpaired t test ( C , D , F ). Two-tailed Student’s paired t test ( H , J , K , L , N , O ). .

Article Snippet: Mouse CTX-I ELISA Kit , Immunodiagnostic Systems , Cat# AC-02F1.

Techniques: Western Blot, Cell Culture, Staining, Isolation, Enzyme-linked Immunosorbent Assay, Two Tailed Test

Journal: EMBO Reports

Article Title: Glutaminolysis provides nucleotides and amino acids to regulate osteoclast differentiation in mice

doi: 10.1038/s44319-024-00255-x

Figure Lengend Snippet: ( A ) Western blot analyses of GLS expression in primary BMM derived from LysMcre;Rosa rtTA (WT) or LysMcre;Gls DOXON mice cultured with 100 ng/mL doxycycline for 48 h ( n = 3). ( B ) TRAP staining of primary BMM isolated from WT or LysMcre;Gls DOXON female mice cultured with RANKL for 2 or 4 days in the presence of doxycycline ( n = 4). Scale bar: 500 μm. ( C ) Quantification of TRAP-positive multinucleated mOC ( n = 4 independent experiments). ( D ) qPCR showing enhanced expression of osteoclast marker genes in LysM;Gls DOXON BMM differentiated for 4 days in the presence of doxycycline ( n = 3 independent experiments). ( E , F ) Representative toluidine blue staining ( E ) and quantification ( F ) showing enhanced resorption pit area in LysM;Gls DOXON BMM differentiated in the presence of doxycycline ( n = 3 independent experiments). Scale bar: 100 μm. ( G , H ) Representative μCT ( G ) and calculated bone volume per tissue volume (BV/TV) ( H ) of trabecular bone in the distal femurs of 2-month-old WT and LysM;Gls DOXON female mice ( n = 9 mice). Scale bar: 200 μm. ( I – J ) Serum ELISA for TRAP 5b ( I ) and CTX-I ( J ) from 2-month-old WT and LysM;Gls DOXON female mice ( n = 9). ( K – L ) Representative TRAP staining ( K ) and quantification of osteoclast number per bone surface (Oc.N/BS) ( L ) from distal femurs from 2-month-old WT and LysM;Gls DOXON female mice ( n = 5). Scale bar: 100 μm. ( M, N ) Representative H&E stain ( M ) and quantification of osteoblast number per bone surface (Ob.N/BS) ( N ) ( n = 5). Scale bar: 100 μm. ( O ) Serum ELISA for P1NP ( n = 9). Data are shown as mean ± SD. Two-tailed Student’s unpaired t test ( C , D , F ). Two-tailed Student’s paired t test ( H , I , J , L , N , O ). .

Article Snippet: Mouse CTX-I ELISA Kit , Immunodiagnostic Systems , Cat# AC-02F1.

Techniques: Western Blot, Expressing, Derivative Assay, Cell Culture, Staining, Isolation, Marker, Enzyme-linked Immunosorbent Assay, Two Tailed Test

Journal: EMBO Reports

Article Title: Glutaminolysis provides nucleotides and amino acids to regulate osteoclast differentiation in mice

doi: 10.1038/s44319-024-00255-x

Figure Lengend Snippet: ( A ) Schematic showing the experimental design. Representative μCT images ( B – F , scale bar: 200 μm), H&E staining ( G – J , scale bar: 100 μm), TRAP staining ( K – O , scale bar: 100 μm), Serum CTX-1 ( P ) and double calcein labeling ( Q – U , scale bar: 50 μm) of distal femur trabecular bone of sham and OVX mice treated with vehicle or CB-839 ( n = 8 mice). ( F ) The calculated BV/TV from µCT images ( n = 8 mice). ( O ) Osteoclast number per bone surface (Oc.N/BS) quantified from TRAP stains ( n = 4 mice). ( P ) Serum ELISA of CTX-I ( n = 8 mice). ( U ) Bone formation rate (BFR) calculated from the double calcein labeling ( n = 4 mice). Data are shown as mean ± SD. Two-way ANOVA ( F , O , P , U ). .

Article Snippet: Mouse CTX-I ELISA Kit , Immunodiagnostic Systems , Cat# AC-02F1.

Techniques: Staining, Labeling, Enzyme-linked Immunosorbent Assay

Journal: EMBO Reports

Article Title: Glutaminolysis provides nucleotides and amino acids to regulate osteoclast differentiation in mice

doi: 10.1038/s44319-024-00255-x

Figure Lengend Snippet: Reagents and tools table

Article Snippet: Mouse CTX-I ELISA Kit , Immunodiagnostic Systems , Cat# AC-02F1.

Techniques: Recombinant, Plasmid Preparation, Sequencing, SYBR Green Assay, Protease Inhibitor, Red Blood Cell Lysis, cDNA Synthesis, Enzyme-linked Immunosorbent Assay, Software

Journal: eLife

Article Title: Csf1 from marrow adipogenic precursors is required for osteoclast formation and hematopoiesis in bone

doi: 10.7554/eLife.82112

Figure Lengend Snippet: ( A ) Representative fluorescent TRAP staining images of femoral long bones from control and Csf1 CKO Adipoq mice at 3 months of age show TRAP+ osteoclasts at different skeletal sites: secondary spongiosa (SS), chondro-osseous junction (COJ), and endosteal surface (Endo.S). TB: trabecular bone; CB: cortical bone. Scale bar = 50 μm. ( B ) Quantification of osteoclast surface (Oc.S) at three skeletal sites. BS: bone surface. L: COJ length. n=5 mice/group. ***, p<0.001 CKO vs control. ( C ) Representative TRAP staining images of osteoclast culture derived from control and Csf1 CKO Adipoq BMMs at 7 days after addition of RANKL and Csf1. Arrows point to mature osteoclasts. Scale bar = 200 μm. ( D ) Quantification of TRAP+ multinucleated cells (>3 nuclei/cell) per field. n=7 mice/group. ( E ) Representative Osterix staining of trabecular bone from control and Csf1 CKO Adipoq femurs. Scale bar = 50 μm. ( F ) Quantification of osteoblast surface (OB.S). BS, bone surface. n=8–12 mice/group. ( G ) Representative double labeling of trabecular bone from control and Csf1 CKO Adipoq femurs. ( H ) Bone formation activity is quantified. MAR: mineral apposition rate; MS: mineralizing surface; BFR: bone formation rate. n=4 mice/group. ( I ) Serum ELISA analysis of bone resorption marker (CTX-1) and formation marker (PINP) in control and CKO mice. n=6–8 mice/group. *, p<0.05 CKO vs control. Figure 4—source data 1. Full dataset for . Figure 4—source data 2. Full dataset for . Figure 4—source data 3. Full dataset for . Figure 4—source data 4. Full dataset for . Figure 4—source data 5. Full dataset for .

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, AFG Scientific, Northbrook, IL, USA) and N-terminal propeptide of type I procollagen (Immunotag Mouse PINP ELISA Kit, G-Bioscience, St. Louis, MO, USA) respectively, according to the manufacturer’s instructions.

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