Journal: Research

Article Title: Visomitin Attenuates Pathological Bone Loss by Reprogramming Osteoclast Metabolism via the STAT3/LDHB Axis

doi: 10.34133/research.0784

Figure Lengend Snippet: Visomitin diminishes the intracellular ROS levels and attenuates osteoclastogenesis. (A) The relative antioxidant capacity of indicated antioxidants at 300 nm was evaluated in the ABTS system ( n = 3). (B and C) Evaluation and quantification of the intracellular ROS levels of BMMs exposed to H 2 O 2 after pretreatment of a range of antioxidants (300 nm) using flow cytometry ( n = 3). (D and E) Detection and quantification of the mean fluorescence intensity (MFI) of DCFH-DA probe in BMMs following treatment with either RANKL or Visomitin; scale bars, 100 μm ( n = 5). (F and G) Detection and quantification of the MFI of MitoSOX probe in BMMs following treatment with either RANKL or Visomitin; scale bars, 50 μm ( n = 5). (H) BMMs were treated with different dosages of Visomitin and subjected to in vitro osteoclast differentiation. Representative images of TRAP staining were shown. Scale bars, 50 μm. (I) Quantification of TRAP + multinuclear cells per well in panel (A) ( n = 3). (J) BMMs were subjected to in vitro osteoclast differentiation and treated with 300 nm Visomitin at specified stages. Representative images of TRAP staining were shown. Scale bars, 50 μm. (K) Quantification of TRAP + multinuclear cells per well in panel (C) ( n = 3). (L) Representative images of wheat germ agglutinin (WGA) staining in osteoclasts treated with or without Visomitin. Scale bars, 10 μm. (M) Quantification of bone pit depth in panel (I) ( n = 12). (N) Representative SEM images of bone slice resorption pits. Scale bars, 10 μm. (O) Quantification of bone resorption pit area ( n = 6). Data are mean ± SD; * P < 0.05, ** P < 0.01, and *** P < 0.001; ns, not significant.

Article Snippet: Staining was then performed with the TRAP Kit (G1492, Solarbio, China).

Techniques: Flow Cytometry, Fluorescence, In Vitro, Staining

Journal: Research

Article Title: Visomitin Attenuates Pathological Bone Loss by Reprogramming Osteoclast Metabolism via the STAT3/LDHB Axis

doi: 10.34133/research.0784

Figure Lengend Snippet: Administration of Visomitin alleviates pathological bone loss in vivo. (A) Representative 3D micro-CT images of the calvaria from mice subjected to either sham or LPS injection, followed by treatment with PBS or Visomitin. Scale bars, 2 mm. (B) Quantification of BV/TV (%) in panel (A) ( n = 5). (C) Representative TRAP and DHE staining of the calvaria from designated groups. Scale bars, 200 μm and 100 μm, respectively. (D to F) Quantification of N.Oc/BS (mm −1 ), Oc.S/BS (%), and relative DHE MFI in panel (C) ( n = 5). (G) Representative 3D micro-CT images of the femurs from mice subjected to either sham or OVX operation, followed by treatment with PBS or Visomitin. Scale bars, 500 and 200 mm, respectively. (H) Quantification of BV/TV (%), Tb.N (mm −1 ), Tb.Th (mm), and Tb.Sp (mm) in panel (G) ( n = 6). (I) Representative TRAP and DHE staining of the femurs from designated groups. Scale bars, 50 μm. (J to L) Quantification of N.Oc/BS (mm −1 ), Oc.S/BS (%), and relative DHE MFI in panel (I) ( n = 6).Data are mean ±SD; * P < 0.05, ** P < 0.01, and *** P < 0.001; ns, not significant.

Article Snippet: Staining was then performed with the TRAP Kit (G1492, Solarbio, China).

Techniques: In Vivo, Micro-CT, Injection, Staining

Journal: Research

Article Title: Visomitin Attenuates Pathological Bone Loss by Reprogramming Osteoclast Metabolism via the STAT3/LDHB Axis

doi: 10.34133/research.0784

Figure Lengend Snippet: Visomitin regulates osteoclastogenesis through the LDHB–lactate axis. (A) The heatmap depicting the gene expression profiles of metabolic pathways. (B) Representative immunoblots depicting the protein levels of LDHA and LDHB following Visomitin treatment ( n = 3). (C) Immunofluorescence staining of LDHB in BMMs subjected to osteoclast differentiation, with or without Visomitin treatment; scale bars, 20 μm. (D) Quantification of the relative LDHB MFI in panel (C) ( n = 6). (E) BMMs infected with either the vector or LDHB-overexpressing adenovirus were differentiated into osteoclasts in the presence or absence of Visomitin treatment. Representative images of TRAP staining were shown. Scale bars, 50 μm. (F) Quantification of TRAP + multinuclear cells per well in panel (E) ( n = 3). (G) The classification of metabolites detected through metabolomics. (H and I) The volcano plot and heatmap illustrating the metabolite affinity profiles derived from metabolomics. (J and K) Enrichment analysis of differential metabolites detected by metabolomics using SMPDB and KEGG databases. Data are mean ± SD; * P < 0.05, ** P < 0.01, and *** P < 0.001; ns, not significant.

Article Snippet: Staining was then performed with the TRAP Kit (G1492, Solarbio, China).

Techniques: Gene Expression, Western Blot, Immunofluorescence, Staining, Infection, Plasmid Preparation, Derivative Assay