Fig 2: Immunoblot analysis of kidney lysates.(A) Blot membranes were incubated with the antibodies against Nrf2, p53 and 8-oxoguanine glycosylase a (OGG1a). The mean integrated optical density is related to actin for Nrf2 (B), p53 (C) and mature 8-oxoguanine glycosylase a (D) levels. Results are given as a mean ± S.E.M. KD4, KD6, KD8 – groups of Eker rats (Tsc2+/-) treated with HFKD for four, six or eight mo., respectively; ST – Eker rats fed with a standard diet; LE ST – wild-type Long Evans rats treated with a standard diet; LE KD – wild-type Long Evans rats treated with a ketogenic diet similarly to the KD6 group. *P < 0.05, **p < 0.01, ***p < 0.001 as compared to ST unless otherwise stated (horizontal line) for ANOVA with a Fisher post hoc test.

Fig 3: Effect of gestational hypoxia compared to normoxia on expression of cellular senescence proteins in the ovary. Data shown as means ± sem. Data are represented as the percentage of the 4-mo normoxia group (assigned baseline value of 100%). Open bars: normoxia (21% oxygen) during gestation; gray bars: hypoxia (13% oxygen) during gestation. P53, left; P16ink, right. ***P < 0.001, *P < 0.05.

Fig 4: Expression of selected proteins in plasma and tumour tissue of treated mice. (A) Plasma of A549 tumour-bearing mice. (B) Plasma of HCT116 tumour-bearing mice. (C) Analysis of HCT116 tumour tissue by ELISA. (D) Analysis of HCT116 tumour tissue by Western blot. PDGF (AA) and CCL2 proteins were determined in plasma obtained from mice bearing (A) A549 and (B) HCT116 tumours and in lysates obtained from HCT116 tumours (C). The levels of VEGF, PDGF, CCL2, FGF1, FGF2 and IL-6 were detected using ELISA according to the manufacturer's protocol. (D) The levels of Ang-4, COX-2, p53, VEGFR2 and eNOS were determined according to the procedure mentioned in the Jess user guide from ProteinSimple. After the detection of targeted proteins by chemiluminescence or NIR fluorescence, the primary and secondary antibodies were removed from capillaries to evaluate the total protein content in a single run. Compass software was used to program in the Jess language and analysis of the results. Quantification was performed by densitometry by calculating the area of targeted proteins and normalizing to the total protein content. Results are expressed as fold change in the expression of proteins when compared to control. N = 6. Statistical analysis: Sidak's multiple comparisons test. *p < 0.05; **p < 0.01