Figure 4

OPN silencing sensitized AML12 hepatocytes to cell death induced by OGD and caused increased NQO1 expression and decreased ATP levels. After silencing of OPN with siRNA in AML12 cells, cell viability (MTT assay) (a) (n=6), LDH release (b) (n=6), cell apoptosis (c) (n=4), cleaved caspase 3 level (d) (n=2), the ATP level (e) (n=4) and gene expression of NQO1 (f) (n=3) were evaluated in the basal state and in response to KCN (2.5, 5 or 10 mM as indicated) after 1 h in DMEM without glucose, followed by a 16- (a, b and f) or 2-h (c, d and e) restoration period in DMEM, 4.5 g/l glucose supplemented with 0.5% BSA. Results relative to either its own control (si Ctr or si OPN) (a and b) or si Ctr (c, d and f) were expressed as means±S.E.M. Data were statistically analyzed using Student’s t-test. *P<0.05