Journal: iScience
Article Title: Mycobacterium tuberculosis specific protein Rv1509 modulates osteoblast and osteoclast differentiation via TLR2 signaling
doi: 10.1016/j.isci.2025.112107
Figure Lengend Snippet: Rv1509 Induces osteoclast differentiation and RANKL secretion by osteoblasts via TLR2 (A) After inducing osteoclast differentiation for four days in the presence of Rv1509 and C29, TRAP staining (scale bar: 400 μm) and F-actin staining (scale bar: 100 μm) were performed, followed by quantitative analysis. The data are shown as mean ± SEM. ( n = 3; ∗∗, p < 0.01; ∗∗∗, p < 0.001 [Student’s t test]). (B) Following a four-day period of osteoclast differentiation stimulated by Rv1509 and C29, RT-qPCR was employed to evaluate the mRNA transcription levels of genes associated with osteoclasts. Additionally, ELISA was performed to measure the expression levels of inflammatory factors. The data are shown as mean ± SEM. ( n = 3; ∗∗, p < 0.01; ∗∗∗, p < 0.001 [Student’s t test]). (C) After 14 days of osteoblast differentiation induced by Rv1509 and C29, ALP staining was performed, followed by Alizarin red staining after 21 days. The data are shown as mean ± SEM. (scale bar: 1 mm; n = 3; ∗, p < 0.05; ∗∗, p < 0.01; ∗∗∗, p < 0.001 [Student’s t test]). (D) The mRNA expression levels of osteoblast-related genes were analyzed using RT-qPCR after 14 days of differentiation in the presence of Rv1509 and C29. The data are shown as mean ± SEM. ( n = 3; ns, p > 0.05; ∗, p < 0.05; ∗∗, p < 0.01; ∗∗∗, p < 0.001 [Student’s t test]). (E) Protein blots showing the representative expression levels of Ctsk, NFATc1, MMP9, and c-Fos in osteoclasts treated with Rv1509 and C29. (F) Protein blots showing representative levels of Col1, Osterix, and Runx2 in osteoblasts after treatment with Rv1509 and C29.
Article Snippet: Rabbit anti-Col1 Ab , ABclonal , Cat# A22089;RRID: AB_3665644.
Techniques: Staining, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Expressing