Figures & data
Figure 1. TNF-α inhibited cell viability and induced apoptosis of HNPC. HNPC were treated with different concentrations of TNF-α (30, 50 and 100 ng/ml), and the cell viability (A) and apoptosis (B,C) was measured using CCK-8 and flow cytometry assay, respectively. **p < .01 compared with control.
Figure 2. TNF-α increased TRIM14 and NF-κBp65 expression of HNPC. HNPC were treated with different concentrations of TNF-α (30, 50 and 100 ng/ml), and the expression of TRIM14 and NF-κBp65 was measured using Western blot and Real-time PCR assay, respectively. **p < .01 compared with control.
Figure 3. TRIM14 down-regulation inhibited apoptosis and related protein expression in HNPC induced by TNF-α. HNPC were treated with 100 ng/ml of TNF-α with or without pLKO.1-shRNA-TRIM14 (shTRIM14) and PDTC treatment. (A) TRIM14 mRNA expression was determined using Real-time PCR assay. (B,C) HNPC apoptosis was determined using flow cytometry assay. (D,E) The level of TRIM14, NF-κBp65, p-NF-κBp65, Bax and Bcl-2 protein was measured by Western blot assay. **p < .01 compared with control. ##p < .01 compared with TNF-α treatment.
Figure 4. TRIM14 up-regulation induced apoptosis and related protein expression in HNPC. HNPC were transfected with pLVX-Puro-TRIM14 expressing vector. (A) TRIM14 mRNA expression was determined using Real-time PCR assay. (B,C) HNPC apoptosis was determined using flow cytometry assay. (D,E) The level of TRIM14, NF-κBp65, p-NF-κBp65, Bax and Bcl-2 protein was determined using Western blot assay. **p < .01 compared with control.
Figure 5. TRIM14 binds to and ubiquitination of PPM1A in vitro. (A,B) The protein expression of PPM1A and PPM1B in TNF-α-induced HNPC was measured by Western blot assay. (C) Co-immunoprecipitation showed that TRIM14 interacts with PPM1A in TNF-α-induced HNPC with TRIM14 down-regulation. (D) HNPC with TRIM14 down-regulation was treated with TNF-α, and PPM1A was immunoprecipitated and immunoblotted. **p < .01 compared with control.
Figure 6. TRIM14, PPM1A, TNF-α and NF-κBp65 mRNA expression in patients with cervical spondylosis. The mRNA expression of TRIM14 (A), PPM1A (B), TNF-α (C) and NF-κBp65 (D) expression in primary human nucleus pulposus tissues in patients with cervical spondylosis (n = 30) and normal subject controls (n = 15) was measured by Real-time PCR. Data are expressed as mean ± SD. **p < .01 compared with control.
