*2.1. Materials*

The key materials and resources used in this study are summarized in Table 1. All other chemicals used were commercially available and of analytical grade. The full-length human ABCC11 WT (NCBI accession: NM\_033151) open reading frame in pcDNA3.1/hygro(−) plasmid [4] and recombinant adenoviruses for the expression of the human ABCC11 WT [10] were constructed in our previous studies; the plasmid/adenovirus vectors and the corresponding control vectors were prepared as a new experimental lot in this study. The plant materials (Table A1) were purchased from local supermarkets in Shizuoka, Japan, between July 2016 and July 2017.




#### **Table 1.** *Cont.*

Purity: ≥98%


**Table 1.** *Cont.*

#### *2.2. Preparation of Plant Extracts*

After the fruits were cleaned, the peels and pulps were carefully separated. The fresh and dried materials (summarized in Table A1) were finely chopped with a knife and ground using a mill (Crush Millser IFM-C20G; Iwatani, Tokyo, Japan), respectively. In the subsequent extraction step, approximately 50 g of the preprocessed plant material were well liquidized in 100 mL of distilled water using a juicer (Crush Millser IFM-C20G; Iwatani) and stirred for 30 min at room temperature. The suspension was centrifuged at 12,000× *g* at 4 ◦C for 10 min to remove the debris. The supernatant was collected and passed through ordinary filter paper. The filtrate was dialyzed against distilled water (500 mL) at 4 ◦C overnight with a dialysis membrane with a molecular weight cut-o ff of 14,000 (Spectrum Chemical Mfg, New Brunswick, NY, USA). The distilled water containing the small molecules that passed the dialysis membrane was lyophilized using FDU-2000 (EYELA, Tokyo, Japan). The freeze-dried extracts were stored at −20 ◦C, dissolved in ultrapure water at 10 mg/mL (10,000 ppm), and subjected to sonication as appropriate before use. Then, 5 μL of the solution were mixed with 20 μL of a transport bu ffer (10 mM Tris/HCl, 250 mM sucrose, and 10 mM MgCl2, and pH 7.4); 1 μL of this clear liquid was used for a vesicle transport assay (total 20 μL/sample), as described below.

## *2.3. Cell Culture*

Human embryonic kidney 293 (HEK293)-derived 293A cells were maintained in Dulbecco's Modified Eagle's Medium (Nacalai Tesque, Kyoto, Japan) supplemented with 10% fetal bovine serum (Biowest, Nuaillé, France), 1% penicillin-streptomycin (Nacalai Tesque), 2 mM L-glutamine (Nacalai Tesque), and 1 × non-essential amino acid (Life Technologies, Tokyo, Japan) at 37 ◦C in a humidified atmosphere of 5% CO2 in air (*v*/*v*), following our previous study [11]. To obtain ABCC11-expressing 293A cells for the plasma membrane vesicles, we performed plasmid transfection using polyethylenimine MAX (1 mg/mL in Milli-Q water, pH 7.0; Polysciences, Warrington, PA, USA) [12] or adenovirus infection [9], as described previously.

#### *2.4. Preparation of ABCC11-Expressing Plasma Membrane Vesicles*

Plasma membrane vesicles were prepared from ABCC11-expressing 293A cells or control cells, as described previously [12], and then rapidly frozen in liquid N2 and stored at −80 ◦C until use. Unless otherwise indicated, the plasma membrane vesicles used in the present study were derived from 293A cells 48 h after the plasmid transfection. The protein concentration of the plasma membrane vesicles was quantified using a BCA Protein Assay Kit (Pierce, Rockford, IL, USA) with bovine serum albumin as a standard according to the manufacturer's protocol.
