Fig 1.
The fruit image and the anthocyanin content in XJ-1~XJ-5 of red-fleshed apple.
Table 1.
Primers sets for qRT-PCR.
Fig 2.
Intracellular reactive oxygen species (ROS) levels assayed with 2’,7’-dichlorofluorescein diacetate (DCFH-DA) fluorescent probe in porcine granulose cells (GCs) treated with Rosup and/or anthocyanins.
A. Blank control (untreated with DCFH-DA probe; untreated with Rosup or anthocyanins); B. Negative control (treated with DCFH-DA probe; untreated with Rosup or anthocyanins); C. Positive control and 0 μM anthocyanins (just treated with 250 μg/mL Rosup); D. 20 μM anthocyanins and Rosup (250 μg/mL); E. 60 μM anthocyanins and Rosup (250 μg/mL); F, 200 μM anthocyanins and Rosup (250 μg/mL).
Fig 3.
Effect of anthocyanin treatment on intracellular ROS levels assayed with 2’,7’-dichlorofluorescein diacetate (DCFH-DA) and dihydroethidium (DHE) fluorescent probe in porcine granulose cells (GCs).
A (A’). Control (untreated with 250 μg/mL Rosup); B (B’). 200 μM anthocyanins; C (C’). 250 μg/mL Rosup; D (D’). 200 μM anthocyanins and 250 μg/mL Rosup; E (E’). Quantitative data for DCFH-DA and DHE. R-A-: no Rosup or anthocyanin treatment (control); R-A+: anthocyanin treatment while no Rosup treatment; R+A-: Rosup treatment while no anthocyanin treatment; R+A+: both Rosup and anthocyanin treatment. The results are expressed as averages ± SE, * indicates P < 0.05, ** indicates P < 0.01.
Fig 4.
The mRNA levels of superoxide dismutase (SOD1), catalase (CAT) and glutathione peroxidase (GPX1) in porcine granulose cells (GCs) treated with anthocyanins and/or Rosup for different times (0–12 h).
A-E. Gene expression in treated cells at 0 h, 3 h, 6 h, 9 h and 12 h. The expression level of β-actin was used as control. R-A-: no Rosup or anthocyanin treatment (control); R-A+: anthocyanin treatment while no Rosup treatment; R+A-: Rosup treatment while no anthocyanin treatment; R+A+: both Rosup and anthocyanin treatment. The relative fold-changes were presented as mean ± SD. Compared to control (R-A-), * indicates P < 0.05, ** indicates P < 0.01.
Fig 5.
The activity of enzymes superoxide dismutase (SOD1), catalase (CAT) and glutathione peroxidase (GPX1) and the content of glutathione (GSH) in porcine granulose cells (GCs).
A. The activity of SOD1; B. The activity of CAT; C. The activity of GPX1; D. GSH content. All values were normalized to protein level and presented as relative fold changes in comparison to untreated control. Data are present as mean ± SE. R-A-: no Rosup or anthocyanin treatment (control); R-A+: anthocyanin treatment while no Rosup treatment; R+A-: Rosup treatment while no anthocyanin treatment; R+A+: both Rosup and anthocyanin treatment. * indicates P < 0.05, ** indicates P < 0.01.
Fig 6.
Data for TdT-mediated dUTP-biotin nick end labeling (TUNEL) assay.
A. Untreated cells (No Rosup or anthocyanin treatment); B. 200 μM anthocyanin-treated cells; C. 250 μg/mL Rosup-treated cells; D. 200 μM anthocyanin and 250 μg/mL Rosup-treated cells; E. The quantitative data. R-A-: no Rosup or anthocyanin treatment (control); R-A+: anthocyanin treatment while no Rosup treatment; R+A-: Rosup treatment while no anthocyanin treatment; R+A+: both Rosup and anthocyanin treatment. * indicates P < 0.05, ** indicates P < 0.01.
Fig 7.
Protein levels of Bcl-2 and Bax in porcine granulose cells (GCs) by Western blotting.
A. Images for Bcl-2 and Bax in porcine GCs in different treatments. Actin was used as the loading control. B. The relative protein level (Bcl-2/Bax). R-A-: no Rosup or anthocyanin treatment (control); R-A+: anthocyanin treatment while no Rosup treatment; R+A-: Rosup treatment while no anthocyanin treatment; R+A+: both Rosup and anthocyanin treatment. Compared to the control group, * indicates P < 0.05, ** indicates P < 0.01.