Siegr1

When the optimal concentration of DEX on cells was determined, HCCLM3 cells were treated with 10 nmol/L DEX for 24 h and transfected with miR-130a mimic, mimic NC, miR-130a inhibitor or inhibitor NC. At the same time, cells were also transfected with miR-130a mimic, mimic NC, miR-130a inhibitor, inhibitor NC, si-EGR1, si-NC, oe-EGR1 or oe-NC, respectively. MiR-130a mimic, inhibitor, and the corresponding NCs were compounded by Shanghai Sangon Biotechnology Co. Ltd. (Shanghai, China) while oe-EGR1, si-EGR1, and the corresponding NCs were constructed by Ribobio (Guangzhou, China). Lipofectamin 2000 reagent (Invitrogen Inc., Carlsbad, CA, USA) was adopted for cell transfection.

Human proximal tubular epithelial (HK-2) cells were cultured as described previously [19 (link)]. Depending on the experiments, HK-2 cells were treated with recombinant human TGF-β1 (10 ng/mL; Gibco, New York, NY, USA), pENTER-Egr1 plasmid (2 μg; Vigene Biosciences, Shandong, China), and small interfering RNA targeting Egr1 (siEgr1, 50 nM, RiboBio, Guangzhou, China). Transfections were performed using Lipofectamine™ 3000 reagent (Invitrogen, USA) following the manufacturer's instructions when cells were cultured to approximately 50–60% confluence in 12-well plates. Cells were harvested 48 h after transfection. TGF-β1 (10 ng/ml) was added to the culture medium 24 h before cell collection.