Mir 143 mimic

Porcine TGF-β1 was purchased from R&D Systems (Minneapolis, MN, USA). Antibodies against PKA, ERK1/2, and p-ERK1/2 were obtained from Sangon Biotech (Shanghai, China), those against AKT and p-AKT were from Cell Signaling Technology (Beverly, MA, USA), and those against FSHR and GAPDH as well as anti-rabbit and anti-mouse IgG were from Santa Cruz Biotechnology (Santa Cruz, CA, USA). DMEM-F-12, fetal bovine serum (FBS), and phosphate-buffered saline were obtained from Life Technologies (Carlsbad, CA, USA). Lipofectamine 2000 was purchased from Invitrogen (Carlsbad, CA, USA). Protease and phosphatase inhibitors were obtained from Roche (Basel, Switzerland). miR-143 mimics, miR-143 inhibitors, and siRNAs were all synthesized by GenePharma (Shanghai, China) (Supplementary Table S1).

MDA-MB-231 and MCF-7 cells were selected to explore the impacts of SBF2-AS1 on BC cells, which were separated into 7 groups: the blank group (the cells without any transfection); the si-negative control (NC) group (cells were transfected with silenced SBF2-AS1 NC vector); the si-SBF2-AS1 group (cells were transfected with silenced SBF2-AS1 vector); the Mimics NC group (cells were introduced with miR-143 Mimics NC); the miR-143 mimics group (cells were introduced with miR-143 mimics); the overexpressed (oe)-SBF2-AS1 + Mimics NC group (cells were transfected with oe-SBF2-AS1 vector and miR-143 Mimics NC); the oe-SBF2-AS1 + miR-143 mimics group (cells were transfected with oe-SBF2-AS1 vector and miR-143 mimics). Mimics NC, miR-143 mimics, si-NC, si-SBF2-AS1 and oe-SBF2-AS1 were all designed and synthesized by Shanghai GenePharma Co., Ltd. (Shanghai, China). The medium was changed to the serum-free medium that without penicillin and streptomycin in the transfection. According to the instructions of Lipofectamine™ 2000 reagent, the silcenced or overexpressed plasmids, and mimics or its NC were mixed and placed for 20 min. The medium was replaced by normal complete medium after the cells were transfected for 6–8 h.