FIG. 8.

FANCJ depletion sensitizes cells to TMS. (A) Whole-cell extracts from HeLa or U2OS cells transfected with control or FANCJ siRNA were probed by Western blotting with antibodies against FANCJ or actin (as a loading control). (B) The proliferation of control or FANCJ siRNA-treated HeLa or U2OS cells was assessed by Coulter counting of the total populations of cells at the indicated time points. (C) WST-1 assay of the proliferation of control cells and cells transfected with FANCJ siRNA. OD₄₅₀, optical density at 450 nm. (D) Colorimetric BrdU cell proliferation ELISA of control or FANCJ siRNA-treated HeLa or U2OS cells. (E) Effect of FANCJ depletion on TMS-induced apoptosis. HeLa or U2OS cells were siRNA treated for 72 h, and cells were incubated with 5 μM TMS for 2 h, washed, and allowed to recover in complete medium for 12 h. Enrichment with the cytoplasmic histone-associated DNA fragments is indicative of apoptosis among cells with the indicated siRNA oligonucleotides. The data shown are the means and SD of results from three independent experiments.