Ab242295

RV gene expression of different inflammatory and oxidative stress markers were assessed at mRNA level by standard qPCR. Macrophage infiltatrion in the RV was assessed by cluster of differentiation 68 (CD68) staining, as decribed previously [34] . Advanced oxidation protein products (AOPP) (ab242295, Abcam, Cambridge, United Kingdom) and anti-oxidant capacity assay (ab65329, Abcam, Cambridge, United Kingdom) were performed in RV tissue to the manufacturer's instruction.
Plasma levels of growth differentiation factor 15 (GDF-15) were measured by ELISA (MGD150, R&D, USA). AOPP were assessed in plasma as well (ab242295, Abcam, Cambridge, United Kingdom).

An advanced oxidation protein products (AOPP) assay, which uses Chloramine-T as a marker of oxidative protein damage, was conducted on the plasma. The AOPP assay kit (ab242295; Abcam), was used in accordance with manufacturer's instructions. In summary, treated and non-treated plasma samples were diluted in PBS (1:10). 200 µL diluted plasma (n = 3), and standards of Chloramine-T (0-100-µM), were transferred to a 96-well plate and 10 µL of Chloramine Reaction Initiator was added to all wells. The plate was shaken for 5 min (120 rpm, room temperature) and 20 µL Stop Solution was added. Absorbance at 340 nm was measured, and AOPP levels expressed as µM of Chloramine-T equivalents, with values corrected for dilution factor.

Freshly collected blood samples from the mice were stood at room temperature for 2 h and centrifuged (1000g, 4°C) for 10 min. Serum hormones follicle-stimulating hormone (FSH), luteinizing hormone (LH), and estradiol (E2) were detected using the ELISA kit (CSB-E06871m, CSB-E12770m, CSB-E07280m, Cusabio, Wuhan, China) in strict accordance with the instructions. The absorbance was measured at 450 nm. The ovarian and oocytes sample lysis fluid were diluted to the optimal concentration to meet the standard curve for detection. Biomarkers related to oxidative stress including total antioxidant capacity (T-AOC), 8-hydroxy-2 deoxyguanosine (8-OH-dG), advanced oxidation protein products (AOPP), molondialdehyde (MDA), superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) were detected by ELISA assay kit (A015-1-2, A001-3-2, A005-1-2, Jiancheng Bioengineering Institute, Nanjing, China; ab201734, ab242295, ab118970, Abcam, USA) following the manufacturer's instructions. The absorbance values were measured using a microplate reader (VersaMax, Molecular Devices, USA).