Mirzip control

Lentiviral vector (pLV-miR-143) for overexpression of miR-143 or an unspecific control (pLV-control) sequence was purchased from Biosettia (Biosettia Inc., San Diego, CA, USA). Both of them contained RFP reporter and showed puromycin resistance. Lentiviral vector for knockdown of miR-143 (miRZip-143) and the corresponding unspecific control (miRZip-control) vector were purchased from System Biosciences (System Biosciences, LLC, Palo Alto, CA, USA). Both of them contained GFP reporter and showed puromycin resistance. Pseudoviral particles were produced according to the manufacturer’s instructions using psPAX2, pMD2G in 293TN cells. Target cells were infected for 24 h with PEG-concentrated (System Biosciences) viral particles followed by puromycin selection or FACS sorting. Purity of transduced cells was analyzed using flow cytometry for expression of fluorescent markers and was > 90%, 7 days after selection or FACS sorting. Stable transduced cells were maintained in media supplemented with 1 μg/mL puromycin.

The miR-210 antagomir and scramble control were purchased from RiboBio Co., Guangzhou, China. The miR-210 agomir and agomir control were synthesized by GenePharma, Shanghai, China. The miR-Zip Control and miR-Zip-210 plasmids were purchased from System Bioscience, Palo Alto, CA, USA. The pLenti and pLenti-STMN1- Myc-DDKvectors were obtained from OriGene, Rockville, MD, USA. The pCMVΔR8.9, pDVsVg, and pLKO.1-shLuc vectors and the shRNA against STMN1 were obtained from the National RNAi Core Facility of Taiwan for gene silencing. All clones were verified by direct sequencing.