Figure 7
Noxa levels influence the sensitivity of GSC-ECL to BH3-mimetics. (a) Cells were transfected with either NT siRNA or Noxa specific siRNA and exposed 48 h to ABT-263 (1 μM) or WEHI-539 (1 μM), combined or not with chemotherapeutic agents. Percentage of cell death (PI+ cells) was determined by flow cytometry in GSC-ECLs. Bar charts show the mean ± S.E.M. of three independent experiments. Student's t-test was conducted to detect significant differences *P < 0.05, **P < 0.01, ***P < 0.001. (b) mRNA levels of Noxa were analyzed by RT-qPCR after 48 h of incubation with the indicated treatments. Bar charts show the mean ± S.E.M. of three independent experiments. Student's t-test was conducted to detect significant differences between untreated and treated cells. *P < 0.05, **P < 0.01, ***P < 0.001. (c) Protein levels of Noxa were analyzed by Western blot after 48 h of incubation with the indicated treatments. Actin was used as loading control. Full-length images are shown in Supplementary Figs. S22 and S23. Bar graphs representing densitometric quantifcation of bands are depicted in Supplementary Fig. S1. (d) G03 and G09 cells were transfected with either NT plus Noxa siRNAs or Bcl-2 plus Bcl-w plus Noxa siRNAs and exposed 48 h to WEHI-539 (1 μM). Percentage of cell death (PI+ cells) was determined by flow cytometry in GSC-ECLs. Bar charts show the mean ± S.E.M. of three independent experiments. For each cell line, the Y-axis scale was set based on the range of the values in the corresponding data set.
