Journal: Frontiers in Aging Neuroscience

Article Title: DNA Demethylation Upregulated Nrf2 Expression in Alzheimer’s Disease Cellular Model

doi: 10.3389/fnagi.2015.00244

Figure Lengend Snippet: Upregulation of mRNA and protein levels of Nrf2 by 5-Aza treatment in N2a/APPswe cells . N2a/APPswe cells were treated with 0 μM 5-Aza (control), 0.1% DMSO, 5-Aza (3 and 5 μM) for 72 h. The expressions of Nrf2 and β-actin mRNA were measured by Q-PCR using total RNA from N2a/APPswe cells. β-Actin was assessed as a loading control (A) . The protein expressions of total Nrf2 (T-Nrf2) and β-actin were measured by Western blot analysis using lysates from N2a/APPswe, normalized to β-actin expression (B) . All data were represented as a mean ± SD of three independent experiments. * P < 0.05 and ** P < 0.01.

Article Snippet: Results have shown that Nrf2 promoter methylation decreased to 39 out of 50 (78%) in the first five CpG sites after treated with 5 μM 5-Aza (chi-square test, P < 0.05) (Figure B), indicating that the first five CpG sites of Nrf2 promoter have been demethylated by 5-Aza treatment.

Techniques: Control, Western Blot, Expressing

Journal: Frontiers in Aging Neuroscience

Article Title: DNA Demethylation Upregulated Nrf2 Expression in Alzheimer’s Disease Cellular Model

doi: 10.3389/fnagi.2015.00244

Figure Lengend Snippet: The first five CpG sites are specifically demethylated by 5-Aza treatment . N2a/APPswe cells were treated with 0.1% DMSO, 5 μM 5-Aza for 72 h. The methylation patterns of the first five CpG sites of promoter Nrf2 gene in N2a/APPswe cells were performed using bisulfite genomic sequencing (BGS) as described in Section “ .” Black circles indicate methylated CpGs, and open circles indicate unmethylated CpGs. The first five CpGs were hypermethylated in N2a/APPswe cells, which were treated with 0.1% DMSO (98% methylated) (A) and cells treated with 5 μM 5-Aza for 72 h (78% methylated) (chi-square test, P < 0.05) (B) .

Article Snippet: Results have shown that Nrf2 promoter methylation decreased to 39 out of 50 (78%) in the first five CpG sites after treated with 5 μM 5-Aza (chi-square test, P < 0.05) (Figure B), indicating that the first five CpG sites of Nrf2 promoter have been demethylated by 5-Aza treatment.

Techniques: Methylation, Genomic Sequencing

Journal: Frontiers in Aging Neuroscience

Article Title: DNA Demethylation Upregulated Nrf2 Expression in Alzheimer’s Disease Cellular Model

doi: 10.3389/fnagi.2015.00244

Figure Lengend Snippet: 5-Aza promotes the nuclear translocation of Nrf2 in N2a/APPswe cells . N2a/APPswe cells were treated with 0 μM 5-Aza (control), 0.1% DMSO, 5-Aza (3 and 5 μM) for 72 h. The protein expressions of N-Nrf2 (nucleus protein of Nrf2) and Lamin B were measured by Western blot using nuclear protein from N2a/APPswe cells. Lamin B was assessed as a loading control (A) . Immunofluorescence staining was used to observe the distribution of Nrf2 (B) . N2a/APPswe cells were treated with 5 μM 5-Aza for 72 h and labeled with anti-Nrf2 antibody (red) and DAPI (blue), APPswe were labeled with GFP (green). Scale bar 50 μm. All data were represented as a mean ± SD of three independent experiments. ** P < 0.01.

Article Snippet: Results have shown that Nrf2 promoter methylation decreased to 39 out of 50 (78%) in the first five CpG sites after treated with 5 μM 5-Aza (chi-square test, P < 0.05) (Figure B), indicating that the first five CpG sites of Nrf2 promoter have been demethylated by 5-Aza treatment.

Techniques: Translocation Assay, Control, Western Blot, Immunofluorescence, Staining, Labeling

Journal: Frontiers in Aging Neuroscience

Article Title: DNA Demethylation Upregulated Nrf2 Expression in Alzheimer’s Disease Cellular Model

doi: 10.3389/fnagi.2015.00244

Figure Lengend Snippet: 5-Aza increases mRNA and protein expression levels of Nrf2 downstream gene NQO1 . N2a/APPswe cells were treated with 0 μM 5-Aza (control), 0.1% DMSO, 5-Aza (3 and 5 μM) for 72 h. The protein expressions of NQO1 and β-actin were measured by Western blot analysis using total protein from N2a/APPswe. β-Actin was assessed as a loading control. All data were represented as a mean ± SD of three independent experiments. * P < 0.05 and ** P < 0.01.

Article Snippet: Results have shown that Nrf2 promoter methylation decreased to 39 out of 50 (78%) in the first five CpG sites after treated with 5 μM 5-Aza (chi-square test, P < 0.05) (Figure B), indicating that the first five CpG sites of Nrf2 promoter have been demethylated by 5-Aza treatment.

Techniques: Expressing, Control, Western Blot