A549 cells (Korean Cell Line Bank, Seoul, Korea; 10185) were cultured in RPMI 1640 medium (Welgene, Gyeongsan, Korea) with 10% heat-inactivated fetal bovine serum (FBS; Gibco, Thermo Fisher Scientific, Waltham, MA) and 1% penicillin/streptomycin (Gibco). Platinum-A (Plat-A) (Cell Biolabs, San Diego, CA; RV-102) cells were grown in Dulbecco’s Modified Eagle’s Medium (Welgene) supplemented with 10% FBS (Gibco) and 1% penicillin/streptomycin and selected by 5 μg/ml puromycin (Sigma-Aldrich) and 10 μg/mL blasticidin (Sigma-Aldrich) prior to produce retroviruses. All cells were incubated at 37°C in with 5% CO2.
Plat a
Manufactured by Cell Biolabs
The Plat-A is a cell line that stably expresses the MLV (Murine Leukemia Virus) Gag, Pol, and envelope (Env) proteins. It is designed for the production of recombinant retroviruses.
Automatically generated - may contain errors
Lab products found in correlation
Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
Penicillin streptomycin, by Merck Group (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Blasticidin, by Merck Group (1 mentions)
Puromycin, by Merck Group (1 mentions)
Rpmi 1640 medium, by Welgene (1 mentions)
Dulbecco s modified eagle s medium, by Welgene (1 mentions)
2 protocols using plat a
1
Culturing A549 and Plat-A Cells
2
Stable Overexpression of XLOC_000647
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For stable overexpression of XLOC_000647, the full-length XLOC_000647 cDNA was synthesized by GENEWIZ (Suzhou, China) and cloned into pBABE retroviral vector (RTV-001-PURO, Cell Biolabs, CA, USA), named as XLOC_000647-pBABE, which were confirmed by sequencing (BioSune, Shanghai, China). After transfection of Plat-A (Cell Biolabs) cells for 48 h, the retrovirus supernatants were collected. Retroviral particles were concentrated by using Retro-Concentin Virus Precipitation Solution (ExCell Bio, Shanghai, China) according to the manufacturer’s guidelines. Then cancer cells were infected with virus and polybrene overnight. Positive clones were screened with puromycin for 2–3 weeks to establish XLOC_000647 stable expression cell lines and corresponding negative control for further study.
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