Figure 6.

Influence of the NC on gene and protein expression of a subset of selected susceptibility-related factors. (A) mRNA expression of a selected subset of genes involved in DDR and DNA-repair, apoptosis, oxidative stress, stress response and transport were analyzed after 24 h mono-treatment of BxPC3 cells with SA (3 µM), NQ (2 µM) and IQ (8 µM) via quantitative RT-PCR. Data shown are the mean values from one representative experiment performed in triplicate. Pooled RNA samples isolated from three independent experiments (n = 3) were used for analysis. Relative mRNA expression in corresponding untreated cells (Con) was set to 1.0 (marked with continuous line). mRNA expression levels were normalized to β-actin. Only changes in mRNA expression of ≥2.0-fold and ≤0.5-fold were considered as biologically relevant (marked with dashed lines). Nd, not detectable. (B,C) Protein expression of repair- (B) and DDR-/apoptosis (C)-related factors was analyzed 24 h after mono-treatment of BxPC3 cells with SA, NQ and IQ. After incubation period of 24 h, BxPC3 cells were harvested for Western blot analyses. Relative protein expression in untreated cells, which were used as control (Con), was set to 1.0. Expression of β-actin, talin-1 or GAPDH was used as protein loading control. Shown are results of a representative experiment. BAX, BCL-2 associated X protein; BCL-2, B-cell lymphoma 2; BRCA1, BReast CAncer Gene 1; CHK1/2, checkpoint kinase 1/2; ERCC1, excision-repair cross-complementing 1; FASL, Fas ligand 1; FASR, Fas receptor; GADD45A, growth arrest and DNA damage-inducible alpha; Mre11, meiotic recombination gene 11; RAD51, RAD51 recombinase; IQ, 5-epi-ilimaquinone; NQ, 5-epi-nakijiquinone Q; SA, secalonic acid F.