Fluorochrome

The subpopulation of thymic lymphocytes was assessed by flow cytometry using antibodies to CD3, CD4, and CD8 conjugated with fluorochromes (Invitrogen, Carlsbad, CA, USA). The sample preparation procedure was carried out according to standard protocols; an FC500 flow cytometer (Beckman Coulter, Brea, CA, USA) was used for the study. The percentage of CD3-positive lymphocytes was determined, including single-positive T-helpers (CD3⁺CD4⁺CD8⁻) and T-cytotoxic (CD3⁺CD4⁻CD8⁺) cells, as well as double-positive (CD3⁻CD4⁺CD8⁺) and negative (CD3⁻CD4⁻CD8⁻) cells representing lymphoblasts.

Chemicals were of analytical grade and, unless otherwise stated, purchased from Sigma-Aldrich (Steinheim, Germany). Fluorochromes were obtained from Life Technologies (Darmstadt, Germany), and semen extender media were obtained from Minitüb (Tiefenbach, Germany).

Ethacrynic acid (EA; Sigma-Aldrich, Saint Louis, MO, USA) was reconstituted in dimethyl sulfoxide (DMSO) to a stock solution of 64 mM. Fluorochromes used for flow cytometry analysis were purchased from Life Technologies (ThermoFisher; Carlsbad, CA, USA) and reconstituted in DMSO, except for propidium iodide (PI) and peanut agglutinin conjugated with fluorescein isothiocyanate (PNA-FITC), which were diluted in phosphate-buffered saline 1× (PBS). The antibody against GSTM3 (ref. ARP53561_P050), as well as its specific blocking peptide (ref. AAP53561), were purchased from Aviva Systems Biology (San Diego, CA, USA). Secondary antibodies for immunoblotting analysis were goat anti-rabbit and rabbit anti-mouse conjugated with horseradish peroxidase (ref. P0448 and ref. P0260; Dako, Derkman A/S; Denmark, respectively), whereas that for immunofluorescence analysis was an anti-rabbit antibody conjugated with Alexa Fluor 488 (ref. A32731; ThermoFisher).

Chemicals were of analytical grade and, unless otherwise stated, purchased from Sigma-Aldrich (Steinheim, Germany). Fluorochromes were obtained from Life Technologies (Darmstadt, Germany), and semen extender media were obtained from Minitüb (Tiefenbach, Germany).

Unless stated otherwise, all reagents were of analytic grade and were purchased from Sigma-Aldrich (Saint-Louis, MO, USA). Fluorochromes were acquired from Molecular Probes (Thermofisher Scientific; Waltham, MA, USA).

All chemicals were purchased from Sigma-Aldrich (St. Louis, MO, USA) unless otherwise stated. Fluorochromes were purchased from Molecular Probes (Thermo Fisher Scientific; Waltham, MA, USA). All reagents are listed in the Table S1.