Fig. 8

Targeting the Prominin-2/BTB and CNC homolog 1 (BACH1) axis to ameliorate rats’ lumbar intervertebral disc degeneration (IVDD) process. Six male Sprague–Dawley (SD) rats aged 4 weeks were used in each group. Establishment of rats’ lumbar IVDD model via puncture surgery. A 24 h after puncture surgery, the rats’ spinal tissue was observed via the EGFP immunostaining under a confocal microscope (n = 6). White Bars = 100 μm. B The intensity of EGFP fluorescence signals was represented (green staining). C 24 h after puncture surgery, the rats’ spinal tissue was harvested and EGFP mRNA levels were determined by RT-PCR (n = 6). D and E and F At pre-surgery, four weeks post-surgery, and eight weeks post-surgery, representative radiographic images of rat lumbar were recorded by X-ray (n = 6). Black arrows: The lumbar intervertebral disc heights. G and H The disc height index (DHI %) was calculated at four weeks and eight weeks post-surgery. I At four weeks post-surgery and eight weeks post-surgery, representative safranin O-fast green (S–O) staining images were shown by performing S–O staining (n = 6). White Bars = 500 μm. J At four weeks post-surgery and eight weeks post-surgery, hematoxylin and eosin (H&E) staining images were shown by performing H&E staining (n = 6). White Bars = 500 μm. K and M Quantitative analysis of S–O staining analysis. L and N Quantitative analysis of H&E staining analysis. O At eight weeks post-surgery, degenerated IVDs were collected, and the proteins were extracted. MMP-9, ADAMTS5, and MMP-13 were evaluated by Western Blot (n = 5). P-R: Quantitative analysis of the protein levels of MMP-9, ADAMTS5, and MMP-13. All data are representative of at least five separate experiments. All these experiments were repeated by at least five times. Analysis: ANOVA followed by a post hoc analysis (B, C, G, H, K, L, M, N, P, Q and R); *P < 0.05, **P < 0.01, ***P < 0.001