Journal: Journal of Alzheimer's Disease

Article Title: Dihydropyridine Derivatives Modulate Heat Shock Responses and have a Neuroprotective Effect in a Transgenic Mouse Model of Alzheimer’s Disease

doi: 10.3233/jad-150860

Figure Lengend Snippet: Fig. 3. Effect of heat and LA1011 or LA1044 treatment on Hsp70, Hsp27 and Hsp40 levels in SH-SY5Y human neuroblastoma cell line measured by western blotting (A, B) and quantitative densitometric analysis (C, D). Cells were untreated or treated with LA1011 (5 M) or LA1044 (40 M) at 37◦C or 42◦C. Box plots represent quantitative data normalized to GAPDH. All data represent the mean ± SD (n = 6) and p < 0.05 was considered statistically significant. HS = heat shock at 42◦C, 1 h. ∗p < 0.05, when data are compared to LA compound (–), HS (–) sample. #p < 0.05, when data are compared to LA compound (–), HS (+) sample. One way ANOVA followed by a Tukey post hoc test was used for statistical comparisons.

Article Snippet: The effect of drug candidates on the stress response was tested with the SH-SY5Y (ATCC® CRL-2266TM) human neuroblastoma cell line stably transfected with a plasmid containing the Hsp70 promoter fused to yellow fluorescent protein (YFP) (pEYFP-N1, Clontech), a kind gift of Lea Sistonen (Åbo Akademi University, Turku, Finland) (SH-SY5Y-pHsp70-YFP).

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,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