invivomab anti mouse rankl cd254 clone ik22 5  (Bio X Cell)

Journal: JBMR Plus

Article Title: Neutralization of Receptor activator of nuclear factor-κB ligand reduces fibrosis and promotes osteoblast differentiation in a mouse model of fibrous dysplasia driven by somatic expression of Gnas R201H

doi: 10.1093/jbmrpl/ziaf145

Figure Lengend Snippet: Anti-RANKL treatment of Gnas R201H ;R26 Tom mice shifts both WT and mutant cells to a more mature osteoblast phenotype, demonstrated by increased OCN and decreased SMA expression. (A-F) Representative confocal images of femur sections from 3-wk-old Gnas R201H ;R26 Tom treated with IgG2a or aRANKL. (A) The proximal metaphyseal region of the femur from 3-wk-old IgG2a and aRANKL treated Gnas R201H ;R26 Tom mutant mice shows endogenous TdTomato (TOM + ) signal in Gnas R201H mutant cells with DAPI nuclear staining. Scale bar = 100 μm. (B and C) Cells in the ROI extending 500 μm below the growth plate were quantified to determine (B) TOM + cells per tissue area (TOM + /T.Ar (/μm 2 )) and (C) TOM + cells per trabecular bone area (TOM + /B.Ar (/μm 2 )) and TOM + cells per bone marrow area (TOM + /bone marrow area (/μm 2 )) from IgG2a and aRANKL treated Gnas R201H ;R26 Tom mice. (D-F) Images of Gnas R201H ;R26 Tom femur sections immunostained for SMA, OSX, or OCN, with DAPI nuclear stain. Endogenous tdTomato signal identifies Gnas R201H mutant cells. (G-L) Cells in the ROI extending 500 μm below the growth plate were quantified to determine (G). Percentage of TOM + Gnas R201H mutant cells that co-expressed SMA (SMA + TOM + /TOM + (%)); (H) Total SMA + cells in the ROI tissue area (SMA + /T.Ar (/μm 2 )); (I) Percentage of TOM + Gnas R201H mutant cells that co-expressed OSX (OSX + TOM + /TOM + (%)); (J) Total OSX + cells in the ROI tissue area (OSX + /T.Ar (/μm 2 )); (K) Percentage of TOM + Gnas R201H mutant cells that co-expressed OCN (OCN + TOM + /TOM + (%)); and (L) Total OCN + cells per ROI tissue area (OCN + /T.Ar (/μm 2 )). Statistical analysis was performed using non-parametric Mann–Whitney t -test, where p < .05 was considered significant.

Article Snippet: Frozen femoral sections were stained for immunofluorescence for SMA (Abcam, ab5694), Ki67 (Cell Signaling, 12202S), osterix (OSX; Abcam, ab22552), osteocalcin (OCN; Thermo Fisher 23 418-1-AP), RANKL (BE0191; BioXCell), and RUNX2 (Abcam, ab192256).

Techniques: Mutagenesis, Expressing, Staining, MANN-WHITNEY

Journal: JCI Insight

Article Title: Hydroxyapatite microspheres induce durable pleurodesis and are rapidly cleared by pleural osteoclasts

doi: 10.1172/jci.insight.192981

Figure Lengend Snippet: ( A and B ) Ctsk –/– mice and control littermates were treated with intrapleural HAM and sacrificed on day 14, and pleural surface hydroxyproline ( A ) and adhesion scores ( B ) were determined as in Figure 1. ( C ) In separate experiments, mice were treated with intrapleural HAM and treated with control IgG or anti-RANKL mAb 3 times weekly over 28 days. μCT was performed, and HAM voxel density was estimated by quantitative methods as outlined in Figure 7. Data are shown as mean ± SD. Comparisons were by unpaired t test for 2 groups and by 1-way ANOVA followed by Tukey’s multiple-comparison test. * P < 0.05, ** P < 0.01, and *** P < 0.001

Article Snippet: C57BL/6J mice were treated with i.p. anti-mouse RANKL mAb (Bio-X-Cell, catalog BE0191, clone IK 22-5) or a Rat IgG2a isotype Control IgG (Bio-X-Cell, catalog BE0089, clone 2A-3) as reported , before being sacrificed 28 days after challenge, followed by assessments of particle density by μCT.

Techniques: Control, Comparison