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. 2017 Jan 27;25(3):715–727. doi: 10.1016/j.ymthe.2016.12.020

Figure 6.

Figure 6

miR-483-5p Directly Targets Timp2 in Chondrocytes In Vitro and in Mice

(A) miR-483-5p aligned with the 3′ UTR of Timp2 mRNA. (B and C) ATDC5 cells co-transfected with a reporter carrying a mutant or WT Timp2 3 ′UTR along with miR-483-5p mimics or its control were analyzed by luciferase assay. (D) Western blot and real-time PCR analysis of Timp2 expression in ATDC5 cells transfected with miR-483-5p mimics or inhibitor. (E) Immunofluorescence analysis showed Timp2 expression was significantly lower in cartilage from OA patients than normal controls. Scale bar, 20 μm. (F and G) Immunofluorescence analysis of Timp2-positive cells in the tibial plateau of DMM OA mice after intraarticular injection of LV3-miR-483-5p (F) or antago-miR-483-5p (G) (n = 10). Scale bar, 40 μm. (H) Immunofluorescence analysis of Timp2-positive cells in the tibial plateau of TG483 and control mice 5 weeks after DMM surgery. (I) Immunofluorescence analysis of Timp2-positive cells in the tibial plateau of TG483 and control mice treated with Dox for 12 months. Red arrows indicate positively stained cells. Error bars represent the mean ± SEM. *p < 0.05; **p < 0.01; ns, non-significant (independent-sample t test for two groups).