Figure 2
From: Clusterin regulates β-amyloid toxicity via Dickkopf-1-driven induction of the wnt–PCP–JNK pathway
Aβ-induced gene expression is dependent on p53 and is necessary and sufficient for neurotoxicity. (a) Rat cortical neurons were treated with 10 μM pifithrin-α for 18 h as indicated and subsequently with Aβ1-42(olig) (3 μM, 3 h). The expression levels of DKK1 (Dickkopf-1), EGR1 (early growth response-1) and FOS (FBJ murine osteosarcoma viral oncogene homologue) were determined by qRT-PCR. (b) Neurons were treated as in (b) using 10 μM PRIMA-1 and then with Aβ and qRT-PCR performed. (c) Rat neurons were treated o/n at 7 d.i.c. with control or Pen1 small interfering RNA (siRNA) to DKK1 (160 nM), and then with 3 μM Aβ1-42(olig) for 24 h and cytotoxicity assayed by the live/dead assay. Healthy cells are labelled green and dead cells are red. Scale bar=10 μM. (d) Neurons were treated as in (c) and cell survival determined at 24, 48 and 72 h by the nuclear morphology assay. d.i.c., days in culture; o/n, over night; qRT-PCR, quantitative reverse transcription polymerase chain reaction.
