A rescue factor abolishing neuronal cell death by a wide spectrum of familial Alzheimer's disease genes and Abeta -- Data Supplement - Supplemental Data -- Proceedings of the National Academy of Sciences

Supplementary material for Hashimoto et al. (2001) Proc. Natl. Acad. Sci. USA 98 (11), 6336–6341. (10.1073/pnas.101133498)

Supplemental Figure 6 Fig. 6.
Effects of some of the death trap (DT) clones on neuronal cell death by familial Alzheimer’s disease (FAD) genes. (A) The deduced nucleotide sequence of the Humanin (HN)-encoding DNA. No. 951-1025 encodes HN. This sequence is the deduced sequence comprised from each DT clone. It should be noted that some of the obtained clones had A or C at T⁹³⁵ and G at A¹⁰²⁴ (four different clones with T⁹³⁵/A¹⁰²⁴, C⁹³⁵/A¹⁰²⁴, T⁹³⁵/G¹⁰²⁴, and A⁹³⁵/G¹⁰²⁴ were noted, while the majority was with T⁹³⁵/A¹⁰²⁴); all other sequences were mutually identical. Considering that the DNA library used was constructed from a single individual, these HN-encoding clones might be derived from different genes. (B) Effects of DT63 and DT171 cDNAs on F11/EcR cell death by ecdysone (EcD)-induced expression of V642I-APP. F11/EcR cells were transfected with pIND-V642I-APP with pEF-BOS, DT63, or DT171 cDNA and were treated with (hatched column) or without (open column) 20 µM EcD, as described in Methods. Cell mortality was similarly measured 72 hr after the onset of EcD treatment. All values with error bars indicate means ± SD of three independent transfections. No T, no transfection. (C) Effect of DT63 on F11 cell death induced by expression of FAD genes. F11 cells were transfected with pcDNA or pcDNA coding for V642I-APP, M146L-PS-1, or N141I-PS-2 with pEF-BOS (vec) or DT63 for 72 hr. Cell mortality was similarly measured. (D) Effects of DT29, DT44, and DT171 cDNAs on F11 cell death by transfected FAD genes. As in A, F11 cells were transfected with pcDNA or pcDNA coding for V642I-APP, M146L-PS-1, or N141I-PS-2 with pEF-BOS (pBOS) or DT clone coding pEF-BOS for 72 hr, and cell mortality was similarly measured. DT29 and DT44 are indicated in Fig. 1. As these experiments were performed simultaneously, the basal death rates (no transfection, pcDNA+pBOS) were the same. Similar experiments were performed at least three times.