Generating Stable Cell Lines for IL-32γ Studies

Cells were transiently transfected with control vector (pcDNA3.1( + )–6×Myc), IL-32γ vector, or ITGAV vector per well, using WelFect-EX PLUS transfection reagent in OPTI-MEM, according to the manufacturer’s specification (WelGENE, Seoul, Korea)^{7 (link)}. To generate stable cell lines (control vector and IL-32γ-CD133+ A549 cells), transfected CD133+ A549 cells were cultured in G418 containing growth medium (600 μg/ml) for 4 weeks. G418-resistant colonies were expanded and used.

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Lee Y.S., Kim K.C., Mongre R.K., Kim J.Y., Kim Y.R., Choi D.Y., Song S., Yun J., Han S.B., Yoon D.Y, & Hong J.T. (2019). IL-32γ suppresses lung cancer stem cell growth via inhibition of ITGAV-mediated STAT5 pathway. Cell Death & Disease, 10(7), 506.

Publication 2019

WelFect-EX PLUS transfection reagent OPTI-MEM

A549 cells Cell lines Cells Cultured medium G418 Transfection Vector

Corresponding Organization : Konkuk University

Other organizations : Chungbuk National University, Yeungnam University

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Variable analysis

independent variables

Control vector (pcDNA3.1(+)–6×Myc)
IL-32γ vector
ITGAV vector

dependent variables

Not explicitly mentioned

control variables

Cell line (CD133+ A549 cells)
Transfection reagent (WelFect-EX PLUS)
Growth medium (with G418, 600 μg/ml)

controls

Positive control: Cells transfected with control vector (pcDNA3.1(+)–6×Myc)
Negative control: Not explicitly mentioned

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