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. Author manuscript; available in PMC: 2018 Jan 25.
Published in final edited form as: Nat Med. 2017 Apr 24;23(6):775–781. doi: 10.1038/nm.4324

Figure 3.

Figure 3

Clearance of SnCs slows the development of naturally occurring OA and post-traumatic OA in aged mice. (a) Study design for clearance of naturally occurring p16INK4a-positive SnCs in female INK-ATTAC mice. AP20187 (AP) was administrated intraperitoneally (i.p.) twice a week starting at 12 months of age. (b) Representative images of safranin O and methyl green staining from vehicle-treated (−AP; n = 6) and AP20187-treated (+AP; n = 4) mice. F, femur; T, tibia. Scale bars, 100 μm. (c) OARSI grade for vehicle-treated (n = 6 mice aged 20, 21, 25, 28, 28 and 32 months) and AP20187-treated (n = 4 mice aged 21, 28, 28 and 34 months) mice. (d) Representative whole-body luminescence images on day 28 after ACLT surgery (no surgery, n = 8; Veh treated, n = 7; UBX0101 treated, n = 8) (left) and quantification of luminescence (in arbitrary units, A.U.) at the indicated times after the surgery of male p16-3MR mice aged 19–20 months treated as indicated in Figure 2a (right). Scale bars, 2 cm (no surgery, n = 8; Veh treated, n = 7; UBX treated, n = 8). (e) Quantification of mRNA levels for Cdkn2a, Cdkn1a, Mmp13, Col2a1 and Acan normalized to Actb levels in joints 28 d after ACLT (n = 3 for all groups). (f) Representative images of safranin O and methyl green staining and immunostaining for p16INK4a (brown, arrows; no surgery, n = 4; Veh treated, n = 3; UBX0101 treated, n = 4), Ki-67 (no surgery and Veh treated, n = 3; UBX0101 treated, n = 4) and HMGB1 (brown, arrowheads; no surgery, n = 5; Veh treated and UBX0101 treated, n = 3) on articular cartilage. HC, hyaline cartilage; CC, calcified cartilage. Scale bars, 100 μm. (g) OARSI grade for no-surgery, vehicle-treated and UBX0101-treated ACLT mice (no surgery, n = 5; Veh treated, n = 6; UBX treated, n = 7). (h) The percentage of weight placed on the operated limb versus the contralateral control (left) and the response time of mice after placement onto a 55 °C platform (right) (no surgery, n = 6; Veh treated, n = 10; UBX treated, n = 11). All data are expressed as means, and each data point represents an individual mouse. One-way ANOVA with Tukey’s multiple-comparisons test was used for statistical analysis in e; a two-tailed t-test (unpaired) was used for c, d, g and h. *P < 0.05, **P < 0.01, ***P < 0.001.