Anti mir 128

The antibodies and reagents used in this study are listed in Table S1. All miRNA mimics, small interfering RNA duplexes, miR‐128 inhibitor (anti‐miR‐128) and its control (anti‐NC) were purchased from GenePharma (Shanghai, China). The negative control RNA duplex (NC) was nonhomologous to any human genome sequence. The expression vectors, firefly luciferase reporter plasmids of 3′ UTRs, and miR‐128 promoter reporters were constructed as described in the Materials and Methods S1. All constructs were verified by sequencing. The sequences of all RNA oligoribonucleotides and primers are listed in Table S2.

miRNA overexpression or knockdown was achieved by transfecting cells with a miRNA mimic or inhibitor. Synthetic RNA molecules, including pre-miR-128, anti-miR-128, and scrambled positive control RNA (pre-miR-control and anti-miR-control), were purchased from GenePharma (Shanghai, China). MCF-7/MDA-231 cells were seeded in 6-well plates and transfected with Lipofectamine 2000 (Invitrogen) on the following day when the cells were approximately 70% confluent. For overexpression of miRNAs, 100 pmol of pre-miR-128 were used. For knockdown of miRNAs, 100 pmol of anti-miR-128 were used. After 6 h, the medium was changed to DMEM/L15 that was supplemented with 2% FBS.
A mammalian expression plasmid (pReceiver-M02-HIC1) was purchased from GeneCopoeia (Germantown, MD, USA). The small interfering RNA (siRNA) (sequence: CCUAGUCUCCUCUAUCGCUTT) targeting human HIC1 was synthesized by GenePharma (Shanghai, China). Total RNA or protein was isolated 24 or 48 h after the transfection.