Figure 4

GADD153 increased Bak expression level. SH-SY5Y cells transfected with pcDNA3-GADD 153 or treated CdCl₂ (25 μM, 12 hr). Total cellular protein was extracted and separated by SDS-PAGE. Western blot analysis was performed using anti-GADD 153 (A). Luciferase activitiy was measured 48 hr after co-transfected pcDNA3-GADD 153 and serial deleted Bak promoter region (B). SH-SY5Y cells were treated with 25 μM Cd for 12 h. Cells were cross-linking Histon-DNA complex by adding 37% formaldehyde for 30 min. Histon-DNA complex sheared between 200 and 1000 bp. ChIP products using specific antibodies directed against pre-immune IgG or GADD 153 were amplified by PCR. Input was sonicated chromatin prior to immunoprecipitation (C). Data are from at least three independent experiments. (D) Scheme of the proposed pathways mediating Cd-induced apoptosis.