Fig. 3: RCAN1.4 the ECM synthesis and remodeling by regulating the calcineurin/NFAT signaling pathway.

a Relative SOX9 and MMP13 gene expression in rat NP cells treated with or without FK506 (a calcineurin inhibitor) and stimulated with hypoxia or DMOG treatment. b Western blot analysis of rat NP cells treated with or without FK506 and stimulated with hypoxia or DMOG treatment. The quantified results are shown below. c Immunofluorescence analysis of SOX9 and d type II collagen in rat NP cells treated with or without FK506 and stimulated with hypoxia or DMOG treatment for 48 h. Results of the quantification of SOX9 and type II collagen expression are shown on the right. e Western blot analysis of rat NP cells treated with vector or RCAN1.4 plasmid and stimulated with hypoxia or DMOG treatment. f Immunofluorescence demonstrated that RCAN1.4 overexpression suppressed the nuclear translocation of NFATc1 in rat NP cells under hypoxia. g Relative expression of SOX9 and MMP13 in rat NP cells treated with or without FK506 and stimulated with negative control siRNA or 100 nM RCAN1.4 siRNA. All experiments were performed at least three times. Scale bar: 100 μm. *P < 0.05, **P < 0.01, ***P < 0.005.