Figure 5. Down-regulation of C/EBPβ via over-expression of miR-155 results in decreased SNX27 levels in Down syndrome brains.
(a) Western blot analysis of SNX27, C/EBPβ, APP and GluR1 in the cortex of age-matched controls (Normal) and Down syndrome individuals (DS). Signal intensity of the immunoblots was calculated and normalized to β-actin; n = 7. P values were calculated using a nonparametric t test, *P < 0.05.
(b) Western blot analysis of SNX27 expression in the cortex of human with sporadic Alzheimer’s disease and control brains. Signal intensity of the immunoblots was normalized to β-actin; n = 8. P values were calculated using a nonparametric t test, *P < 0.05.
(c) Quantitative RT-PCR analyses of SNX27 mRNA levels in the cortex of Down syndrome individuals and normal controls. Data represent mean ± s.e.m., n = 7. P values were calculated using a nonparametric t test, *P < 0.05.
(d) Western blot analyses of C/EBPβ and SNX27 protein levels in MC-IXC cells transfected with miR-155 or control mimic. Data represent mean ± s.e.m., n = 3. P values were calculated using two-tailed Student’s t test, **P < 0.01.
(e) Quantitative RT-PCR analyses of SNX27 and C/EBPβ mRNA levels in MC-IXC cells transfected with miR-155 or control mimic. Data represent mean ± s.e.m., n = 3. P values were calculated using two-tailed Student’s t test, *P < 0.05, **P < 0.01.
(f) Western blot analysis for SNX27 expression in MC-IXC cells transfected with LAP2 or control. Data represent mean ± s.e.m., n = 3. P values were calculated using two-tailed Student’s t test, *P < 0.05.
(g) Western blot analysis of Snx27 expression in the cortex and hippocampus from Cebpb+/− and Cebpb−/− mice. Data represent mean ± s.e.m., n = 3. P values were calculated using two-tailed Student’s t test, *P < 0.05.
(h) Firefly luciferase reporter assay for SNX27 promoter activity. Data represent mean ± s.e.m., n = 4. P values were calculated using two-tailed Student’s t test, *P < 0.05.
(i) Binding of C/EBPβ to the SNX27 promoter as determined by a Chromatin IP assay.