The Journal of Neuroscience, June 7, 2006, ():
Presenilin-Dependent 
-Secretase-Mediated Control of p53-Associated Cell Death in Alzheimer's Disease
J. Neurosci. Alves da Costa et al.
26: 6377
Supplemental data
Files in this Data Supplement:
- supplemental material -
Supplemental Data
- supplemental material -
Supplemental References
- supplemental material
-
Fig.S1: Influence of overexpressed and endogenous PS2 on p53. p53 activity (PG13-luciferase, A,C), transactivation of human (hPP-luciferase, A) or murine (mPP-luciferase, C) p53 promoter and p53 expression (B,D) were measured as described in the Methods in HEK293 overexpressing empty pcDNA3 vector (Mock) or PS2 cDNA (A,B) or in wild-type (PS+/+) or PS2-deficient (PS2-/-, C,D) fibroblasts. Bars correspond to the ratios of luciferase/β-galactosidase activities and are the means ± S.E.M of 8 independent experiments. **p<0.001, ***p<0.0001.
- supplemental material
-
Fig.S2: Wild-type and N141I-PS2 activate caspase 3 in a p53-dependent manner. Mock-transfected (Mock), wild-type PS2 (PS2WT) and N141I-PS2 (N141I)-expressing HEK293 cells were assessed for their p53 activity (A), p53 promoter transactivation (B) or p53 mRNA levels (C) as described in the Methods. In D and E, p19Arf-/- (D) and p19Arf-/-p53-/- (E) fibroblasts were transiently transfected with empty cDNA (CT), wild-type PS2 (PS2WT) and N141I (N141I)-PS2. Forty eight hours after transfection, cells were treated for 4 hours with (black bars) or without (white bars) staurosporine (1μM) then caspase 3 activity was measured as described in the Methods. Bars are the means ± S.E.M of 3 independent experiments. *p<0.05; **p<0.01; ***p<0.001. ns, not statistically significant.
- supplemental material
-
Fig.S3: Influence of overexpressed and endogenous PS1 on p53. p53 activity (PG13-luciferase, A,C), transactivation of human (hPP-luciferase, A) or murine (mPP-luciferase, C) p53 promoter and p53 expression (B,D) were measured as described in the Methods in HEK293 overexpressing empty pcDNA3 vector (Mock) or PS1 cDNA (A,B) or in wild-type (PS+/+) or PS1-deficient (PS1-/-, C,D) fibroblasts. Bars correspond to the ratios of luciferase/β-galactosidase activities and are the means ± S.E.M of 8 independent experiments. *p<0.05,**p<0.001, ***p<0.0001.
- supplemental material
-
Fig.S4: Influence of endogenous and overexpressed PS1 on endogenous PS2-like immunoreactivity. Wild-type (PS+/+) and PS1-deficient (PS1-/-) mouse fibroblasts (A,C) or Mock (pcDNA3)- or PS1-transfected HEK293 cells (B,C) were analyzed for their CTF-PS2- and PS1-like immunoreactivities using anti NTF-PS1 or anti-loop PS2 polyclonal antibody (1/1000, gift from Dr G. Thinakaran) as described in the Methods. Four typical independent blots for CTF-PS2 are shown for each cell types. Bars in C represent the densitometric analysis of CTF-PS2 immunoreactivity and are expressed as the percent of control CTF-PS2 recovered in PS+/+ fibroblasts or in HEK Mock-transfected cells. Values are the means ± SEM of 6 independent experiments.
- supplemental material
-
Fig.S5: AICD induces p53-dependent cell death
p19Arf-/- (A) or p19Arf-/-p53-/- (B) were transfected with pcDNA3 or AICDC59 (C59) coding vectors without (-) or with (+) co-transfection with Tip60 and Fe65 cDNA then caspase 3 activity was assessed in presence of staurosporine as described in the Methods. Cells were treated with pifithrin-α (black bars), an inactive pifithrin-α analog (gray bars) or not treated (white bars). Bars are the means ± SEM of 3 to 4 independent determinations. Typical image of p19Arf-/- (C) and p19Arf-/-p53-/- (D) tunel-positive fibroblasts. Cells were transfected with empty (pcDNA3) or AICDC59 (C59) coding vectors, alone or in combination with Tip60 and Fe65 cDNA and treated with 1µM staurosporine before being processed for tunel analysis. (E) shows means of tunel-positive cells ± SEM for 10 independent optical fields in the indicated conditions. ns, non significant,*p<0,01, **p<0,001, ***p<0,0001.
- supplemental material
-
Supplemental Table
