Fig 1: EVs-circHIPK3 derived from MSCs attenuated IL-1ß induced chondrocyte injury. A Overexpressed circHIPK3 was observed in MSC EVs based on qRT-PCR. B With chondrocytes treated with IL-1ß and MSCs-circHIPK3-EVs, the expression of miR-124-3p and MYH9 was detected by qRT-PCR. C MYH9 expression was detected through Western blot post treating chondrocytes with IL-1ß and MSCs-circHIPK3-EVs. D With chondrocytes treated with IL-1ß and MSCs-circHIPK3-EVs, the expression change in cartilage genes (Aggrecan, COL2A1, Runx2, Sox9 and MMP-13) was detected by qRT-PCR. E The expression change in key cartilage genes (Runx2, MMP-13, COL2A1, Aggrecan, and Sox9) was detected through Western blot after chondrocytes were treated with IL-1ß and MSCs-circHIPK3-EVs. F Cell proliferation was detected through EdU experiment with chondrocytes treated with IL-1ß and MSCs-circHIPK3-EVs. G Transwell experiment was conducted for the detection of cell proliferation after treating chondrocytes with IL-1ß and MSCs-circHIPK3-EVs. H A flow cytometer was employed for the detection of cell apoptosis after chondrocytes were treated with IL-1ß and MSCs-circHIPK3-EVs. *P < 0.05, **P < 0.01
Fig 2: MSCs-circHIPK3-EVs inhibited cartilage degradation. We established the OA model, and divided them into five groups: OA model, Normal, OA + MSCs-EVs, OA + MSCs-circHIPK3-EVs and OA + circHIPK3. A HE staining and Mankin score assessment. B the number of PCNA-positive cells using immunohistochemistry. C–D The mRNA and protein levels of Sox9, COL2A1, Aggrecan, Runx2 and MMP-13 were detected by qRT-PCR and western blot
Fig 3: Expression of KAT6B downstream molecules. qPCR was performed to measure the expression of RUNX2 (a), RUNX3 (b), COL2A1 (c), and COL10A1 (d) in primary chondrocytes from control (N = 10) and CS (N = 13) groups. e Western blotting was performed to measure the expression of RUNX2, RUNX3, COL2A1, and COL10A1 in primary chondrocytes from control and CS groups. CS, congenital scoliosis. *p < 0.05
Fig 4: MSCs-circHIPK3-EVs up-regulated COL2A1, Aggrecan, and SOX9 and decreased MMP-13 and Runx2 expression. A After the treatment of OA mouse derived chondrocytes with MSCs-circHIPK3-EVs, the expression changes in these key cartilage genes (COL2A1, Sox9, Runx2, Aggrecan and MMP-13) by qRT-PCR. B After OA mouse derived chondrocytes were treated with MSCs-circHIPK3-EVs, Western blot was performed to detect the expression change in these key cartilage genes (Sox9, COL2A1, Aggrecan, Runx2 and MMP-13). *P < 0.05
Fig 5: sh-MYH9 decreased COL2A1, Aggrecan and Sox9, increased MMP-13 and Runx2. A After the treatment of OA mouse derived chondrocytes with sh-MYH9, MSCs-EVs and MSCs-circHIPK3-EVs, the expression changes in these key cartilage genes (COL2A1, Sox9, Runx2, Aggrecan and MMP-13) by qRT-PCR. B After OA mouse derived chondrocytes were treated with sh-MYH9, MSCs-EVs and MSCs-circHIPK3-EVs, Western blot was performed to detect the expression change in these key cartilage genes (Sox9, COL2A1, Aggrecan, Runx2 and MMP-13). *P < 0.05, **P < 0.01
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