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. 2023 Jun 16;27(16):2340–2353. doi: 10.1111/jcmm.17818

FIGURE 3.

FIGURE 3

Normalize the extracellular matrix (ECM) production of nucleus pulposus (NP) cells by the treatment with melatonin (MLT). NP cells were treated with conditioned medium (CM) in the presence or absence of MLT for 24 h, and followed by different examinations. (A–E) The relative mRNA levels of matrix anabolic and degrading enzymes in NP cells were measured by qPCR analyses (n = 3). (A) The levels of ACNA mRNA were increased in the presence of MLT treatment at different dosages. (B) The levels of COL2A1 mRNA were upregulated in the presence of MLT treatment at different dosages. (C) The levels of MMP13 mRNA were decreased in the presence of MLT treatment at different dosages. (D) The levels of ADAMTS4 mRNA were downregulated in the presence of MLT treatment at different dosages. (E) The levels of ADAMTS5 mRNA were declined in the presence of MLT treatment at different dosages. (F–K) The relative protein levels of matrix anabolic and degrading enzymes in NP cells were measured by Western blot analyses (n = 3). (F) Western blotting showed the different modifications of ECM protein productions in NP cells. (G) Western blotting showed the upregulation of ACNA proteins. (H) Western blotting showed the upregulation of COL2A1 proteins. (I) Western blotting showed the reduction of MMP13 proteins. (J) Western blotting showed the downregulation of ADAMTS4 proteins. (K) Western blotting showed the reduced level of ADAMTS5 proteins. Data are expressed as mean ± standard deviation. The two groups among the five groups are compared by using an independent t‐test. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.