Fig. 4.

NIP1;2 facilitates Al uptake and efflux in yeast. (A) NIP1;2-GFP (green) was colocalized with red CellMask PM Staining (Thermo Fisher Scientific) in a yeast cell carrying pYES2-NIP1;2::GFP. (Scale bar: 1 μm.) Arrows points to the VM. (B) Uptake of Al³⁺, Al-malate (Al-Mal), and Al-citrate (Al-Cit) by NIP1;2. Yeast lines carrying pYES2-GFP or pYES2-NIP1;2-GFP were treated with AlCl₃, Al-Mal, or Al-Cit at pH 4.2 for 1 h. (C) Time-dependent Al-Mal uptake by NIP1;2. Yeast lines were treated with Al-Mal for the indicated times. (D) Time-dependent Al efflux by NIP1;2. Yeast cells carrying pYES2-NIP1;2-GFP were pretreated with Al-Mal for 8 h, then transferred to fresh LPM medium supplemented with 0 or 5 μM AgNO₃ at pH 7.0 for the indicated times. Yeast Al concentration was determined by ICP-MS. The net Al uptake (C) and Al efflux (D) at each time point was the difference between two yeast lines (C) or two treatments (D). Data are mean ± SD of three biological replicates from three independent transformation events. **P < 0.01 between two yeast lines under the indicated treatment conditions.