Journal: Scientific Reports

Article Title: Probable involvement of p11 with interferon alpha induced depression

doi: 10.1038/srep17029

Figure Lengend Snippet: ( A,B ) The p11 protein levels were measured using western blots. The p11 protein levels were normalized against GAPDH. The results demonstrated that IFN-α treatment down-regulated the p11 protein levels in SH-sy5y cells. ( A ) Different doses of hIFN-α-2b (0, 50, 500, 1000, 2000, and 3000 IU/mL) for 24 h produced significant dose-dependent decreases in the p11 protein levels. ( B ) Time-dependent (0, 6, 12, 24, 36, and 48 h) effects in IFN-α (1000 IU/mL) treated cells, with reduced p11 protein levels at 24 h and the lowest levels at 36 h. ( C ) Dose-dependent effect of IFN-α on the p11 mRNA levels in SH-sy5y cells, which were normalized against GAPDH. SH-sy5y cells were treated with hIFN-α-2b (0, 50, 500, 1000, 2000, and 3000 IU/mL) for 16 h, and p11 mRNA levels were measured using real-time PCR. The real-time PCR results demonstrated no effects of hINF-α-2b on the p11 mRNA levels. All results were representative of three separate experiments. The data represented the mean ± S.E. compared with the controls. * P < 0.05 and ** P < 0.01.

Article Snippet: Human neuroblastoma cell line (SH-sy5y cell line, Jiahe Biotech Company, Shanghai, China) was cultured in Dulbecco’s modified Eagle medium (DMEM) containing 10% fetal bovine serum, 50 U/mL penicillin, and 50 mg/mL streptomycin at 37 °C, with 5% CO 2 .

Techniques: Western Blot, Produced, Real-time Polymerase Chain Reaction

Journal: Scientific Reports

Article Title: Probable involvement of p11 with interferon alpha induced depression

doi: 10.1038/srep17029

Figure Lengend Snippet: ( A–C ) IFN-α treatment down-regulated the 5-HTR1b and 5-HTR4 protein levels in a dose-dependent manner. The 5-HTR1b and 5-HTR4 protein levels were determined using western blotting. Cells were treated with different doses of hIFN-α-2b (0, 50, 500, 1000, 2000, and 3000 IU/mL; 0 IU/mL as the control) for 24 h. ( A ) The western blots detected a significant dose-dependent decrease in the 5-HTR1b/4 protein levels after hIFN-α-2b treatment. The 5-HTR1b (B) and 5-HTR4 ( C ) protein levels were normalized against GAPDH. ( D,E ) The real-time PCR results show that IFN-α treatment has no influence on the 5-HTR1b/4 mRNA levels in SH-sy5y cells. Cells were treated with different doses of hIFN-α-2b for 16 h. The values were normalized against GAPDH. ( D ) The 5-HTR1b mRNA levels in hIFN-α-2b treatment groups had no significant difference compared with the controls ( P > 0.05). ( E ) The 5-HTR4 mRNA levels in hIFN-α-2b treatment groups had no significant difference compared with the controls ( P > 0.05). All results were representative of three separate experiments. The data represented the mean ± S.E. compared with the controls. * P < 0.05 and ** P < 0.01.

Article Snippet: Human neuroblastoma cell line (SH-sy5y cell line, Jiahe Biotech Company, Shanghai, China) was cultured in Dulbecco’s modified Eagle medium (DMEM) containing 10% fetal bovine serum, 50 U/mL penicillin, and 50 mg/mL streptomycin at 37 °C, with 5% CO 2 .

Techniques: Western Blot, Control, Real-time Polymerase Chain Reaction

Journal: Scientific Reports

Article Title: Probable involvement of p11 with interferon alpha induced depression

doi: 10.1038/srep17029

Figure Lengend Snippet: ( A ) Western blots showed the 5-HTR1b and 5-HTR4 protein levels in SH-sy5y cells treated with hIFN-α-2b (1000 IU/mL) at various time points (0, 6, 12, 24, 36, and 48 h). ( B,C ) The 5-HTR1b and 5-HTR4 protein levels were normalized against GAPDH. The results showed that the 5-HTR1b ( B ) and 5-HTR4 ( C ) levels declined at 6 h and reached their lowest levels at 24 h. All results were representative of three separate experiments. The data represented the mean ± S.E. compared with the controls. * P < 0.05 and ** P < 0.01.

Article Snippet: Human neuroblastoma cell line (SH-sy5y cell line, Jiahe Biotech Company, Shanghai, China) was cultured in Dulbecco’s modified Eagle medium (DMEM) containing 10% fetal bovine serum, 50 U/mL penicillin, and 50 mg/mL streptomycin at 37 °C, with 5% CO 2 .

Techniques: Western Blot

Journal: Scientific Reports

Article Title: Probable involvement of p11 with interferon alpha induced depression

doi: 10.1038/srep17029

Figure Lengend Snippet: ( A ) SH-sy5y cells were treated with 1000 IU/mL hIFNα-2b, and with an equal volume of PBS as the control. The proteins in cytomembrane of SH-sy5y cells were extracted at 24 h. The p11, 5-HTR1b and 5-HTR4 protein levels were analyzed using western blotting. ( B ) The levels of p11, 5-HTR1b and 5-HTR4 protein in cytomembrane were normalized against Pan-cadherin. Tublin test was performed to eliminate the contamination from cytoplasm to cytomembrane proteins. All results were representative of three separate experiments. The data represented the mean ± S.E. compared with the controls. ** P < 0.01.

Article Snippet: Human neuroblastoma cell line (SH-sy5y cell line, Jiahe Biotech Company, Shanghai, China) was cultured in Dulbecco’s modified Eagle medium (DMEM) containing 10% fetal bovine serum, 50 U/mL penicillin, and 50 mg/mL streptomycin at 37 °C, with 5% CO 2 .

Techniques: Control, Western Blot

Journal: Scientific Reports

Article Title: Probable involvement of p11 with interferon alpha induced depression

doi: 10.1038/srep17029

Figure Lengend Snippet: After plasmid transfection for 24 h, 1000 U/mL hIFNα-2b or PBS was added to the medium. Total proteins were extracted at 48 h. and the p11, 5-HTR1b and 5-HTR4 protein levels were analyzed using western blotting. Total RNA was extracted at 36 h and reverse transcription was applied to produce cDNA. The 5-HTR1b and 5-HTR4 mRNA levels were analyzed using real-time PCR. ( A ) SH-sy5y cells were transfected with p11-pcDNA3.0 containing full-length p11cDNA, with pcDNA3.0 as the control. ( B–D ) The p11 ( B ), 5-HTR1b ( C ), and 5-HTR4 ( D ) protein levels in p11-pcDNA3.0 transfection groups were normalized against GAPDH. ( E ) After PBS treatment, SH-sy5y cells were transfected with p11-miRNA that interfered with the expression of p11, miRNA-control vector as the control. ( F–H ) The p11 ( F ), 5-HTR1b ( G ), and 5-HTR4 ( H ) protein levels in p11-miRNA transfection groups were normalized against GAPDH. ( I,J ) mRNA levels of 5-HTR1b ( I ) and 5-HTR4 ( J ) in SH-sy5y cells in each transfection group was analyzed using real-time PCR and normalized against GAPDH. All results were representative of three separate experiments. The data represented the mean ± S.E. compared with the controls. * P < 0.05, ** P < 0.01 and NS , no significant difference.

Article Snippet: Human neuroblastoma cell line (SH-sy5y cell line, Jiahe Biotech Company, Shanghai, China) was cultured in Dulbecco’s modified Eagle medium (DMEM) containing 10% fetal bovine serum, 50 U/mL penicillin, and 50 mg/mL streptomycin at 37 °C, with 5% CO 2 .

Techniques: Plasmid Preparation, Transfection, Western Blot, Reverse Transcription, Real-time Polymerase Chain Reaction, Control, Expressing