Figure 6.

Effects of Nutlin or silencing of p53 on mRNA levels of TM7SF3, p21, iNOS and CHOP. Human islets (a and c) were transfected for 24 h with 25 nM of p53-siRNA or with a non-targeting siRNA (control). Cells were then treated with nutlin (10 μM) for the indicated periods of time. Alternatively, HepG2^sh–con or HepG2^sh-p53 cells (b, d and g), naive HepG2 (e) or HCT116 cells (f) were treated with Nutlin (10 μM) for 24 h (e) or for the indicated periods of time (a–d, f and g). In parallel, HepG2^sh–control or HepG2^sh-p53 cells were transfected for 48 h with 25 nM of TM7SF3-siRNA or with a non-targeting control sequence. Then the cells were treated with doxorubicin (2 μM; 24 h) (h). RNA was extracted from all cells and quantification of the indicated mRNAs was carried out by qRT-PCR (a–d, f–h). Alternatively, total cell extracts were resolved by SDS-PAGE and immunoblotted with the indicated antibodies (e). Bar graphs are the mean±S.E.M. of at least three independent experiments. *P<0.05, **P<0.01, ***P<0.001 versus non-targeting siRNA assayed under the same conditions