For liposomal transfection complex formation, 15 µl Lipofectamine™ 3000 (cat. no. L3000001, Thermo Fisher Scientific, Inc.) was mixed with 250 µl Opti‑MEM™ reduced serum medium (cat. no. 31985070, Gibco) and incubated at room temperature for 5 min. A total of 15 µl small interfering (si) RNA [ATG7 (Autophagy‑related gene 7) RNA I (cat. no. 6604, Cell Signaling Technology, Inc., sequences not available), NCOA4‑targeting and scramble siRNA (sequences provided in Table I; Tsingke Biotechnology Co., Ltd.)] at a final concentration of 50 nM was separately diluted in 250 µl Opti‑MEM reduced serum medium. Each siRNA diluent was mixed with the Lipofectamine™ 3000‑Opti‑MEM™ mixture, and the transfection complexes were incubated at room temperature for 20 min.
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