miRNA mimics, inhibitors and negative controls were purchased from GenePharma (Shanghai, China). To knock down HIC1, three siRNA sequences targeting different sites of human HIC1 open reading frame (ORF) were designed and synthesized by GenePharma, and the sequence with the best interfering effect was selected (sequence: CCUAGUCUCCUCUAUCGCUTT). For the HIC1 overexpression assay, a mammalian expression plasmid (pReceiver-M02-HIC1) designed to specifically express the full-length ORF of human HIC1 without the miR-23~27~24–responsive 3′-UTR was purchased from GeneCopoeia (Germantown, MD, USA). An empty plasmid served as a negative control. Lipofectamine 2000 (Invitrogen, Carlsbad, CA, USA) was used for transfection according to the instructions. Total RNA and protein were isolated 24 or 48 h after transfection.
Preceiver m02 hic1
Manufactured by GeneCopoeia
Sourced in United States
The PReceiver-M02-HIC1 is a laboratory equipment product manufactured by GeneCopoeia. It is designed for specific research applications, but a detailed description cannot be provided while maintaining an unbiased and factual approach. The core function of this product is not available for disclosure.
Automatically generated - may contain errors
2 protocols using preceiver m02 hic1
1
Manipulating HIC1 Expression in Cells
2
miRNA-mediated regulation of HIC1 expression
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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.
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.
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