Defactinib further protects articular cartilage from degeneration. (A) Safranin O and fast green staining (top). Scale bar, 200 μm. Magnified view (bottom) of Safranin O and fast green staining. Scale bar, 100 μm. (B) International modified Mankin scores of articular cartilage 60 days after surgery. n = 10 per group. (C) H&E staining where double-headed arrows mark calcified cartilage (CC) and hyaline cartilage (HC) thickness. Scale bars, 100 μm. (D) Quantitative analysis of HC/CC ratio in cartilage. n = 10 per group. (E–I) Immunostaining and quantification of MMP-13, ADAMTS 5, Aggrecan, and Col II in articular cartilage 60 days post-operation. n = 10 per group. Scale bar, 100 μm. Sham = sham controls; ACLT + Veh = vehicle-treated ACLT mice; ACLT + Def = defactinib-treated ACLT mice. ∗p < 0.05 and ∗∗p < 0.01 compared to sham. #p < 0.05 and ##p < 0.01 compared to vehicle.