Fig. 3.

Differential effects of downregulation of specific caspases on TFD-induced death of wild-type and caspase-2-null sympathetic neurons. A, Penetratin1-linked antisense oligonucleotides specifically downregulate targeted caspases. PC12 cells were treated with the indicated antisense oligonucleotides (240 nm) for 6 hr. Cells lysates containing equal amounts of protein were subjected to Western blotting using the corresponding antisera. Actin staining confirmed equal loading. These are representative blots; similar results were obtained in two independent experiments. B, C, Sympathetic neurons from P1 wild-type (B) and caspase-2-null (C) mice were cultured for 5 d. Cultures were then washed and treated with anti-NGF in the presence and absence of the indicated antisense oligonucleotides (open squares, control; closed squares, anti-NGF;open diamonds, anti-NGF + V-ACasp1; closed triangles, anti-NGF + V-ACasp2; open circles, anti-NGF + V-ACasp3; closed circles, anti-NGF + V-ACasp6; open triangles, anti-NGF + V-ACasp7;closed inverted triangles, anti-NGF + V-ACasp8;squares with crosses, anti-NGF + V-ACasp9). Cultures were scored daily, and survival is reported relative to the numbers of living neurons in the same cultures before NGF deprivation and is given as the mean ± SEM (n = 3). Error bars are sometimes too small to be visible. This is a representative experiment; similar results were obtained in six independent experiments.