Fig. 7

SAC abrogation triggers cell death through a pathway involving BID, CASP-2 and CASP-3. A Schematic depicting the hypothetical transduction pathways triggered by TTK and AURKB inhibitors in cancer cells. Modified from [28]. B PC9-ER cells, sensitive to SAC abrogation, were treated with the AZD2811 150 nM (AZD L), AZD2811 500 nM (AZD H) or Osimertinib 500 nM (osi) and selected proteins of the pathway presented in (A) were assessed by Western blotting. The images shown are a representative of two different experiments. C Quantification of the immunoblots shown in (B). Bars represent mean ± SEM of two independent experiments. D Parental PC9, seven PC9-derived clones and NCI-H1819 cells were treated with Osimertinib 500 nM (osi) or AZD2811 150 nM (AZD) for 24 h (48 h for NCI-H1819) and selected proteins of the pathway presented in (A) were analyzed by Western blotting. Cells sensitive and resistant to SAC abrogation are indicated in green and blue, respectively. E Quantification of the immunoblots shown in (D). The intensity of the bands was normalized against β–tubulin. F NCI-H1819 cells silenced for BID and PC9-GR3 cells knocked-out for BID or CASP2 were treated with AZD2811 (150 nM) or osimertinib (500 nM) and selected proteins assessed by Western blotting. G Quantification of the bands in the Western blottings in (D) and (F), showing changes in protein levels after AZ2811 treatment. The intensity of the bands was normalized against β–tubulin. H Parental PC9-ER cells and PC9-ER CASP-2 KO were treated with AZD2811 (150 nM) and assessed by Western blotting. I Quantification of the bands corresponding to PC9-ER parental cells in the Western blotting shown in (H). The intensity of the bands was normalized against HSP90