Sigenome risc free sirna

BT549, MDA‐MB‐468 and MDA‐MB‐436 cell lines, which all belong to triple‐negative breast carcinoma cells, were purchased from Cell Bank of the Institute of Basic medicine, Chinese Academy of Medical Sciences (Beijing, China), or obtained as NCI‐ICBP45 kit procured through American Type Culture Collection (ATCC; ATCC Breast Cancer Cell Panel, Manassas, VA, USA). BT549 cells were cultured in RPMI 1640 medium (Boster, Wuhan, China) supplemented with 10% FBS (Gibco, Carlsbad, USA), 1% antibiotics and 0.023 IU·mL⁻¹ bovine insulin (Sigma‐Aldrich, Saint Louis, MO, USA). MDA‐MB‐436 and MDA‐MB‐468 cells were cultured in Dulbecco's modified Eagle's medium (Boster) with 20% or 10% FBS (Gibco) and 1% antibiotics. All cells were kept at 37 °C and 5% CO₂. The short hairpin RNA (shRNA) product of SGLT1 (sh‐SGLT1) with a targeted sequence of ATCTTTCTCTTATTGGCAA and its negative control (sh‐NC) were purchased from GeneChem (Shanghai, China) and transfected into cells according to manufacturer’s instructions. Short interfering RNA (siRNA) oligos against SGLT1 (MU‐007589‐01‐0002) were purchased from Dharmacon (Lafayette, CO, USA). Sequences are available from Dharmacon or upon request. As a negative control, we used siGENOME RISC‐Free siRNA (Dharmacon). Cells were transfected with the indicated siRNA oligos at a final concentration of 35 nm using Dharmafect 2 reagent (Dharmacon).