Cisplatin cp

Cisplatin (CP) (Sigma-Aldrich, St. Louis, MO, U.S.A.) was diluted in saline to make accurate dose for testing. 7.5 mg/kg body weight dose of CP was selected on the basis of previous literature to induce acute renal toxicity [38 ]. Silymarin (100 mg/kg b.w) and AHE (400 mg/kg b.w) were prepared freshly in distilled water before treating.

The mutants gip1, gip2, gip1gip2 and their corresponding genetic background Col-0, WS, and Col-0 x WS have been described previously (Janski et al., 2012 (link); Batzenschlager et al., 2017 (link)). Arabidopsis seedlings were grown in vitro on ½ Murashige and Skoog medium (SERVA Electrophoresis) in presence of 1% sucrose supplemented with 1.2% agar at 20°C under long day conditions (16-h light 70 μmol m^–2 s^–1 of fluorescent lighting/8-h dark). The gip1gip2 mutant was complemented by the expression of a pGIP1::GIP1-GFP construct as previously described (Janski et al., 2012 (link); Batzenschlager et al., 2015 (link)). For sensitivity tests to genotoxic drugs, seeds were initially sowed on ½ MS agar and after 5 days on growth, seedlings were transferred on media supplemented with the genotoxins. Drug concentrations were 10 μM for Bleomycin (BLM) (Laboratoire Thissen, Belgium) and 50 μM for cisplatin (CP) (Sigma, St. Louis, United States). Seedlings were treated during 16 h by 50 μM CP in ½ MS for cytological analyses.

Cisplatin (CP) was purchased from Sigma-Aldrich (USA), whereas sodium citrate was from Egyptian International Pharmaceutical Industries Company, Cairo, Egypt.