Journal: Biomedical Journal

Article Title: Autophagy-induced osteoblast-derived exosomes maintain bone formation and prevent osteoporosis by remodeling gut microbiota-metabolism

doi: 10.1016/j.bj.2025.100870

Figure Lengend Snippet: Auto-exo reduce bone loss and promote osteoblast differentiation in vivo . (A) Schematic diagram of animal experimental design. (B) Quantitative analysis of bone mineral density (BMD) among different groups. (C) HE staining of bone tissues. Scale bar = 100 μm. (D) TRAP staining. Scale bar = 100 μm. (E, F) The expression of osteoblast differentiation-associated genes (RUNX2, OSX, Plzf, Dlx5, and ALP) in OVX mice after Auto-exo treatment were assayed by qRT-PCR (E) and western blot (F). Results are displayed as mean ± SD. ∗ p < 0.05. ∗∗ p < 0.01.

Article Snippet: Following blocking with TBST solution with 5 % skim milk, primary antibodies against CD63 (1:1000, ab216130, Abcam), TSG101 (1:5000, ab125011, Abcam), Calnexin (1:20000, ab92573, Abcam), RUNX2 (1:1000, 20700-1-AP, Proteintech), OSX (1:1000, 28694-1-AP, Proteintech), Plzf (1:5000, 66672-1-Ig, Proteintech), Dlx5 (1:1000, 10592-1-AP, Proteintech), ALP, (1:200, sc-271431, Santa cruz) and β-actin (1:2000, 60008-1-lg, Proteintech) were appended to incubate membranes overnight at 4 °C.

Techniques: In Vivo, Staining, Expressing, Quantitative RT-PCR, Western Blot

Journal: Biomedical Journal

Article Title: Autophagy-induced osteoblast-derived exosomes maintain bone formation and prevent osteoporosis by remodeling gut microbiota-metabolism

doi: 10.1016/j.bj.2025.100870

Figure Lengend Snippet: Bilirubin inhibits osteoblast differentiation of MC3T3-E1 cells. (A) ALP staining. (B) The calcified cells were examined by Alizarin red S staining. (C, D) The expression of osteoblast differentiation-associated genes (RUNX2, OSX, Plzf, Dlx5, and ALP) was assayed by qRT-PCR (C) and western blot (D). NC represents normal control. BL represents low concentration of bilirubin. BM represents medium concentration of bilirubin. BH represents high concentration of bilirubin. Results are displayed as mean ± SD. ∗ p < 0.05, ∗∗ p < 0.01.

Article Snippet: Following blocking with TBST solution with 5 % skim milk, primary antibodies against CD63 (1:1000, ab216130, Abcam), TSG101 (1:5000, ab125011, Abcam), Calnexin (1:20000, ab92573, Abcam), RUNX2 (1:1000, 20700-1-AP, Proteintech), OSX (1:1000, 28694-1-AP, Proteintech), Plzf (1:5000, 66672-1-Ig, Proteintech), Dlx5 (1:1000, 10592-1-AP, Proteintech), ALP, (1:200, sc-271431, Santa cruz) and β-actin (1:2000, 60008-1-lg, Proteintech) were appended to incubate membranes overnight at 4 °C.

Techniques: Staining, Expressing, Quantitative RT-PCR, Western Blot, Control, Concentration Assay

Journal: Biomedical Journal

Article Title: Autophagy-induced osteoblast-derived exosomes maintain bone formation and prevent osteoporosis by remodeling gut microbiota-metabolism

doi: 10.1016/j.bj.2025.100870

Figure Lengend Snippet: Auto-exo accelerated osteoblast differentiation by modulating bilirubin. (A) ALP staining. Scale bar = 200 μm. (B) The calcified cells were detected by Alizarin red S staining. Scale bar = 200 μm. (C) The expression of osteoblast differentiation-associated genes (RUNX2, OSX, Plzf, Dlx5, and ALP) was evaluated by qRT-PCR. Results are displayed as mean ± SD. ∗ p < 0.05, ∗∗ p < 0.01.

Article Snippet: Following blocking with TBST solution with 5 % skim milk, primary antibodies against CD63 (1:1000, ab216130, Abcam), TSG101 (1:5000, ab125011, Abcam), Calnexin (1:20000, ab92573, Abcam), RUNX2 (1:1000, 20700-1-AP, Proteintech), OSX (1:1000, 28694-1-AP, Proteintech), Plzf (1:5000, 66672-1-Ig, Proteintech), Dlx5 (1:1000, 10592-1-AP, Proteintech), ALP, (1:200, sc-271431, Santa cruz) and β-actin (1:2000, 60008-1-lg, Proteintech) were appended to incubate membranes overnight at 4 °C.

Techniques: Staining, Expressing, Quantitative RT-PCR

Journal: Biomedical Journal

Article Title: Autophagy-induced osteoblast-derived exosomes maintain bone formation and prevent osteoporosis by remodeling gut microbiota-metabolism

doi: 10.1016/j.bj.2025.100870

Figure Lengend Snippet: Auto-exo reduce bone loss and promote osteoblast differentiation in vivo . (A) Schematic diagram of animal experimental design. (B) Quantitative analysis of bone mineral density (BMD) among different groups. (C) HE staining of bone tissues. Scale bar = 100 μm. (D) TRAP staining. Scale bar = 100 μm. (E, F) The expression of osteoblast differentiation-associated genes (RUNX2, OSX, Plzf, Dlx5, and ALP) in OVX mice after Auto-exo treatment were assayed by qRT-PCR (E) and western blot (F). Results are displayed as mean ± SD. ∗ p < 0.05. ∗∗ p < 0.01.

Article Snippet: Following blocking with TBST solution with 5 % skim milk, primary antibodies against CD63 (1:1000, ab216130, Abcam), TSG101 (1:5000, ab125011, Abcam), Calnexin (1:20000, ab92573, Abcam), RUNX2 (1:1000, 20700-1-AP, Proteintech), OSX (1:1000, 28694-1-AP, Proteintech), Plzf (1:5000, 66672-1-Ig, Proteintech), Dlx5 (1:1000, 10592-1-AP, Proteintech), ALP, (1:200, sc-271431, Santa cruz) and β-actin (1:2000, 60008-1-lg, Proteintech) were appended to incubate membranes overnight at 4 °C.

Techniques: In Vivo, Staining, Expressing, Quantitative RT-PCR, Western Blot

Journal: Biomedical Journal

Article Title: Autophagy-induced osteoblast-derived exosomes maintain bone formation and prevent osteoporosis by remodeling gut microbiota-metabolism

doi: 10.1016/j.bj.2025.100870

Figure Lengend Snippet: Bilirubin inhibits osteoblast differentiation of MC3T3-E1 cells. (A) ALP staining. (B) The calcified cells were examined by Alizarin red S staining. (C, D) The expression of osteoblast differentiation-associated genes (RUNX2, OSX, Plzf, Dlx5, and ALP) was assayed by qRT-PCR (C) and western blot (D). NC represents normal control. BL represents low concentration of bilirubin. BM represents medium concentration of bilirubin. BH represents high concentration of bilirubin. Results are displayed as mean ± SD. ∗ p < 0.05, ∗∗ p < 0.01.

Article Snippet: Following blocking with TBST solution with 5 % skim milk, primary antibodies against CD63 (1:1000, ab216130, Abcam), TSG101 (1:5000, ab125011, Abcam), Calnexin (1:20000, ab92573, Abcam), RUNX2 (1:1000, 20700-1-AP, Proteintech), OSX (1:1000, 28694-1-AP, Proteintech), Plzf (1:5000, 66672-1-Ig, Proteintech), Dlx5 (1:1000, 10592-1-AP, Proteintech), ALP, (1:200, sc-271431, Santa cruz) and β-actin (1:2000, 60008-1-lg, Proteintech) were appended to incubate membranes overnight at 4 °C.

Techniques: Staining, Expressing, Quantitative RT-PCR, Western Blot, Control, Concentration Assay

Journal: Biomedical Journal

Article Title: Autophagy-induced osteoblast-derived exosomes maintain bone formation and prevent osteoporosis by remodeling gut microbiota-metabolism

doi: 10.1016/j.bj.2025.100870

Figure Lengend Snippet: Auto-exo accelerated osteoblast differentiation by modulating bilirubin. (A) ALP staining. Scale bar = 200 μm. (B) The calcified cells were detected by Alizarin red S staining. Scale bar = 200 μm. (C) The expression of osteoblast differentiation-associated genes (RUNX2, OSX, Plzf, Dlx5, and ALP) was evaluated by qRT-PCR. Results are displayed as mean ± SD. ∗ p < 0.05, ∗∗ p < 0.01.

Article Snippet: Following blocking with TBST solution with 5 % skim milk, primary antibodies against CD63 (1:1000, ab216130, Abcam), TSG101 (1:5000, ab125011, Abcam), Calnexin (1:20000, ab92573, Abcam), RUNX2 (1:1000, 20700-1-AP, Proteintech), OSX (1:1000, 28694-1-AP, Proteintech), Plzf (1:5000, 66672-1-Ig, Proteintech), Dlx5 (1:1000, 10592-1-AP, Proteintech), ALP, (1:200, sc-271431, Santa cruz) and β-actin (1:2000, 60008-1-lg, Proteintech) were appended to incubate membranes overnight at 4 °C.

Techniques: Staining, Expressing, Quantitative RT-PCR