Fig. 5

DMH2 induction of cell death involves the downregulation of pTAK1. a, b Western blot of H1299 and A549 cells treated with DMH2 for 48 h demonstrating an increase in activated caspase-3. (c) Representative images of H1299 cells treated with 1 μ DMH2 for 3 days demonstrating significant cell shrinkage and chromatin condensation. (d, e) H1299 and A549 cells were treated with DMH2 alone or in combination with SB for 3 days and cell death was determined. (f, g) Western blot analysis of cells treated with DMH2 or SB for 3 days. (h) Knockdown of TAK1 was performed in H1299 cells using siRNA. The percentage of dead cells was determined after 3 days. (i) Western blot analysis showing knockdown of TAK1 in H1299 cells. (j, k) DMH1 alone or in combination with SB did not cause significant cell death in H1299 cells after 3 days (k) and did not induce the activation of caspase-3. (l) Western blot analysis demonstrating DMH1 alone or in combination with SB did not show significant regulation of Smad2, pTAK1, or p65. (m) LDN induced little cell death compared to DMH2 and (n) did not downregulate Smad2 or TAK1 by Western blot analysis. All experiments were performed at least 3 times