pCMV6-E2F1, pCMV6-c-myc, and pCMV6-XL5 empty plasmid was purchased from Origene (Rockville, MD). MiR-301a hairpin antagomirs were obtained from GeneCopoeia Inc. (Guangzhou, China). All transfections were conducted using Lipofectamine 2000 according to the manufacturer's instructions (Invitrogen). miR-301a-expressing retroviruses, Cip2a-expressing retroviruses were purchased from GENECHEM (Shanghai, China). The Cip2a short hairpin RNA (shRNA) lentivirus vectors were generated by GeneCopoeia Inc. (Guangzhou, China). The target sequences of Cip2a shRNA was 5'-CUGUGGUUGUGUUUGCACUTT-3'. For selection of TNBC stable cell lines, retroviruses were transduced into BT549 and MDA-MB-231 cells in the presence of polybrene (6 μg/mL, Sigma). Cells were selected with 1 μg/mL puromycin for 7 days.
Pcmv6 xl5 empty plasmid
Manufactured by OriGene
PCMV6-XL5 is an empty plasmid vector designed for cloning and expression in mammalian cells. The plasmid contains a CMV promoter for high-level expression of the inserted gene, and the XL5 backbone for stable maintenance in host cells.
Automatically generated - may contain errors
Lab products found in correlation
Pcmv6 c myc, by OriGene (1 mentions)
Pcmv6 e2f1, by OriGene (1 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (1 mentions)
Polybrene, by Merck Group (1 mentions)
Sch002, by Merck Group (1 mentions)
Lipofectamine 2000 reagent, by Thermo Fisher Scientific (1 mentions)
Gv102, by Genechem (1 mentions)
2 protocols using pcmv6 xl5 empty plasmid
1
Construction of TNBC Cell Lines
2
Lentiviral Manipulation of ALKBH5 and FOXO1
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Lentiviral vectors expressing nontargeting pLKO.1 control shRNA (SCH002), two shRNA constructs targeting ALKBH5-shRNA1 (TRCN0000064783) and -shRNA2 (TRCN0000064787) were obtained from Sigma-Aldrich (St. Louis, MO, USA). Short hairpin sequences against either the FOXO1 or the scrambled short hairpin RNA (shRNA) sequences were cloned into the GFP-labeled lentiviral vector GV102 (GENECHEM). The target sequences selected are: shControl, TTCTCCGAACGTGTCACGT; shFOXO1-1, CAGTCTGTCCGAGATCAGTAA; shFOXO1-2, AGCGGGCTGGAAGAATTCAAT. Expression plasmid for pCMV6-SOD2 (Cat. No RC202330), and pCMV6-XL5 empty plasmid was purchased from Origene (Rockville, MD). For cell transfection, Lipofectamine 2000 reagent (Invitrogen, Carlsbad, CA) was used according to the manufacturer's instructions. After 72 h transfection, stably expressing or knockdown cells were selected in RPMI 1640 medium containing 1 μg/mL puromycin.
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