Fig. 3: iKeap1 ameliorates H2O2-induced death of murine and human osteoblasts.
The primary murine osteoblasts (A–G) or the primary human osteoblasts (H–M) were pretreated (for 2 h) with iKeap1 (2.5/10 μM), followed with or without H2O2 (400 μM) stimulation and osteoblasts were cultured for applied time periods; Cell viability was tested by CCK-8 assays (A and H); Caspase-3 activity was tested (B and I); Expression of apoptosis-associated proteins was tested by western blotting assays (C); Cell apoptosis was examined by nuclear TUNEL staining assays (D and J) and Annexin V FACS (E and K) assays, and results were quantified. Mitochondrial CyPD-ANT1-p53 association and their expressions were shown (F and L), and cell necrosis examined by quantifying medium LDH release (G and M). Expressions of the listed proteins were quantified and normalized to the loading control. Quantified values were mean ± standard deviation (SD, n = 5). “C” stands for the untreated control cells. *P < 0.05 vs. “C” cells. #P < 0.05 vs. cells with H2O2 stimulation but “Veh” pretreatment. Experiments were repeated five times, with similar results obtained.
