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Fig. 1 | Arthritis Research & Therapy

Fig. 1

From: Positive feedback regulation between USP15 and ERK2 inhibits osteoarthritis progression through TGF-β/SMAD2 signaling

Fig. 1

USP15 overexpression can prevent cartilage damage in vivo and in vitro. a ITS induction solution was added to induce cartilage phenotype in ATDC5 cells at 7, 14, and 21 days, and quantitative real-time PCR was used to detect the marker molecules related to cartilage phenotype. b, c Overexpression or knockout of USP15 in ATDC5 cells with TGF-β1 (10 ng/mL). d Immunofluorescence staining performed in ATDC5 cells with or without overexpressing USP15 for Col2a1, Aggrecan, and Col10a1 (scale bars = 50 μm). e The intensities of immunofluorescence of Col2a1, Aggrecan, and Col10a1 in each group were calculated, and the data were presented as the mean ± SD. fh The expression levels of USP15, p-SMAD2, Col2a1, Aggrecan, and MMP13 in the two groups were detected by immunohistochemistry, and the cartilage tissue morphology was detected by HE/Safranin-O fast green staining (n = 4 for control groups in the OA models, n = 4 for AAV-mediated USP15 overexpression groups in the OA models). f Scale bars = 50 μm. g Scale bars = 50 μm. h Scale bars = 200 μm. i The relative expressions of USP15, p-SMAD2, Col2a1, Aggrecan, and MMP13 in each group were calculated via immunohistochemistry, and the hyaline cartilage thickness and OARSI scores were quantified. The data were presented as the mean ± SD. *P < 0.05, **P < 0.01, ***P < 0.001. j Relative gene expressions of Col2a1, Aggrecan, and Sox9 associated with cartilage anabolic metabolism in both groups were detected by quantitative real-time PCR (n = 3 for control groups in the OA models followed by quantitative real-time PCR, n = 3 for AAV-mediated USP15 overexpression groups in the OA models followed by quantitative real-time PCR). The data were presented as the mean ± SD. *P < 0.05, **P < 0.01, ***P < 0.001. All experiments were performed at least three times

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