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FIGURE 8 Limonin exerts the effect of anti-oxidative stress through the NRF2/HO-1 pathway. (A) Frozen liver sections of zebrafish larvae with liver-specific eGFP expression were immunofluorescent stained with NRF2 (n = 6–8 per group). (B) Immunofluorescence staining of HO-1 in zebrafish larvae (n = 6–8 per group). (C–D) Quantitative analysis of the liver mean fluorescence intensity of NRF2 and HO-1. (E) Real-time PCR analysis of NRF2 mRNA levels in zebrafish larvae. The mRNA levels were normalized to β-actin mRNA levels and presented as fold change compared with the control group (n = 3 per group). (F) Western blot analysis of the expression of HO-1 and NRF2. GAPDH was used as an internal control. (G) Relative quantitative protein expression of NRF2 and HO-1. (H) Immunofluorescence staining of HO-1 in LO2 cells. (I) Cellular fluorescence intensity of HO-1. (J) Immunofluorescence staining of NRF2 in LO2 cells. (K) The respective fluorescence intensity of the nucleus, cytoplasm and ratio of NRF2. Data are shown as the mean ± SD from three independent experiments. #p < 0.05, ##p < 0.01, ###p < 0.001 vs control group; *p < 0.05, **p < 0.01, ***p < 0.001 vs TAA group.