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. 2021 Jun 7;9:675486. doi: 10.3389/fcell.2021.675486

FIGURE 6.

FIGURE 6

ISRIB and CPE treatment exerted a protective effect on NP cells in vivo. The bilateral inferior articular processes and supra- and interspinous ligaments were transected to induce lumbar instability of mice. Next, the mice were subcutaneously injected with NS, ISRIB, CPE, or the combination with ISRIB and CPE once a day after surgery and sacrificed after 8 weeks of injection. (A) X-ray of the mice lumbar vertebrae, and DHI of the L3–L4 level (scale bar = 2 mm). (B) Representative micrographs of slices stained with Safranin O (scale bar = 100 μm), Col-II, and Col-X (scale bar = 50 μm), and (C) quantitation of percentage of total positive area or cells (%). Data are presented as means ± SD (n = 6). *p < 0.05; **p < 0.01; ***p < 0.001; the orange column indicates control mice without lumbar instability surgery, the green column indicates mice injected with NS after lumbar instability surgery, the red column indicates mice injected with ISRIB after lumbar instability surgery, the blue column indicates mice injected with CPE after surgery, and the gray column indicates mice injected with ISRIB and CPE after lumbar instability surgery. CPE, cyclopamine; NP, nucleus pulposus; NS, normal saline; DHI, disc height index; Col-II, collagen II; Col-X, collagen X.