Fig 1: Expression of SPRY4 in healthy and degenerated human cartilage.a Safranin-O staining (upper panel) and SPRY4 IHC staining (lower panel) in healthy and degenerated human cartilage. Scale bar, 100 μm. b Chondrogenic marker gene mRNA expression levels (top), hypertrophic marker gene mRNA expression levels (middle), and SPRY4 mRNA expression levels (bottom) in Healthy chondrocytes (HCs) and Degenerated chondrocytes (DCs). The data represent the mean ± SEM (n = 3; ACAN, SOX9, COL2, and MMP13: unpaired t-test; RUNX2 and COL10: unpaired t-test with Welch’s correction; MMP13: Mann Whitney test).
Fig 2: Effect of SPRY4 suppression on the OA-related biological properties of HCs.a mRNA expression analysis of various chondrogenic, hypertrophic, and ECM protease markers in siCON-treated and siSPRY4-treated HCs. The data represent the mean ± SEM (n = 3, unpaired t-test with Welch’s correction). b Western blot protein analyses of various chondrogenic, hypertrophic, and ECM protease markers in siCON-treated and siSPRY4-treated HCs. c Alcian blue and Safranin-O staining of siCON-treated and siSPRY4-treated HCs. Scare bar, 100 μm.
Fig 3: Effect of SPRY4 overexpression on OA-related biological properties of DCs.a mRNA expression analysis of various chondrogenic, hypertrophic, and ECM protease markers in Lenti-CON-treated and Lenti-SPRY4-treated DCs. The data represent the mean ± SEM (n = 3; ACAN and MMP13: unpaired t-test; SOX9, COL2, RUNX2, COL10, and ADAMT5: unpaired t-test with Welch’s correction). b Western blot protein analyses of various chondrogenic, hypertrophic, and ECM protease markers in Lenti-CON-treated and Lenti-SPRY4-treated DCs. c Alcian blue and Safranin-O staining of Lenti-CON-treated and Lenti-SPRY4-treated DCs. Scale bars, 200 μm.
Fig 4: Effect of SPRY4 overexpression on hypertrophy in DCs.a CCK-8 cell proliferation assay for the control lentivirus (Lenti-CON)-treated and SPRY4 lentivirus (Lenti-SPRY4)-treated DCs by time (n = 3, unpaired t-test with Welch’s correction). b Microscopic images of Lenti-CON-treated and lenti-SPRY4-treated DCs (n = 5, unpaired t-test with Welch’s correction) Scale bars, 200 μm. The data represent the mean ± SEM (c) DAPI staining of Lenti-CON-treated and Lenti-SPRY4-treated DCs (n = 5, unpaired t-test with Welch’s correction) Scale bars, 100 μm. d ROS staining of Lenti-CON-treated and Lenti-SPRY4-treated DCs (n = 5, unpaired t-test) Scale bar, 200 μm. e β-gal staining of Lenti-CON-treated and Lenti-SPRY4-treated DCs (n = 5, Mann Whitney test) Scale bar, 200 μm. All data represent the mean ± SEM. Abbreviations: CCK-8, cell counting kit-8; DAPI, 4′,6-diamidino-2-phenylindole; ROS, reactive oxygen species; β-gal, β-galactosidase.
Fig 5: Effect of SPRY4 suppression on chondrocyte hypertrophy in HCs.a CCK-8 cell proliferation assay of siCON-treated and siSPRY4-treated HCs by time (n = 3, unpaired t-test with Welch’s correction). b Microscopic images of siCON-treated and siSPRY4-treated HCs (n = 5, unpaired t-test) Scale bars, 100 μm. c DAPI staining of siCON-treated and siSPRY4-treated HCs (n = 5, unpaired t-test with Welch’s correction) Scale bars, 100 μm. d ROS staining of siCON-treated and siSPRY4-treated HCs (n = 5, unpaired t-test with Welch’s correction) Scale bars, 250 μm. e β-gal staining of siCON-treated and siSPRY4-treated HCs (n = 4, unpaired t-test) Scale bars, 250 μm. All data represent the mean ± SEM. Abbreviations: CCK-8, cell counting kit-8; DAPI, 4′,6-diamidino-2-phenylindole; ROS, reactive oxygen species; β-gal, β-galactosidase.
Supplier Page from Abcam for Anti-Sprouty 4/Spry-4 antibody