Quantifying Apoptosis Markers in Cell Lines

Total RNA was extracted from YCC-2 and SNU-668 cells treated with G. thunbergii using RNAiso Plus reagent (TaKaRa Bio, Japan), and cDNA was synthesized using cDNA Master Mix (ToYoBo, Japan) according to the manufacturer’s protocol. Reverse transcription PCR was performed as described previously (Kim et al. 2018 (link)) using the following PCR primer sequences: Bcl-2, 5ʹ-GGATGCCTTTGTGGAAAACCCTGT-3ʹ (forward) and 5ʹ-AGCCTGCAGCTTTGTTTCAT-3ʹ (reverse); Bax, 5ʹ-TTTGCTTCAGGGTTTCATCC-3ʹ (forward) and 5ʹ-CAGTTGAAGTTGCCGTCAGA-3ʹ (reverse); caspase-3, 5ʹ-TTTTTCAGAGGGGATCGTTG-3ʹ (forward) and 5ʹ-CGGCCTCCACTGGTATTTTA-3ʹ (reverse); PARP, 5ʹ-TGGAACATCAAGGACGAGCT-3ʹ (forward) and 5ʹ-GCATCGCTCTTGAAGACCAG-3ʹ (reverse); GAPDH, 5ʹ-GGCTGCTTTTAACTCTGGTA-3ʹ (forward) and 5ʹ-ACTTGATTTTGGAGGGATCT-3ʹ (reverse). PCR products were used for 1% agarose gel electrophoresis with RedSafe Nucleic Acid Staining Solution (iNtRON Biotechnology).

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Lee H., Kim W., Kang H.G., Kim W.J., Lee S.C, & Kim S.J. (2019). Geranium thunbergii extract-induced G1 phase cell cycle arrest and apoptosis in gastric cancer cells. Animal Cells and Systems, 24(1), 26-33.

Publication 2019

RNAiso Plus reagent cDNA Master Mix RedSafe Nucleic Acid Staining Solution

Agarose gel electrophoresis Bcl 2 Caspase 3 Cdna Cells Gapdh Intron Nucleic acid Primer Reverse transcription

Corresponding Organization :

Other organizations : Chosun University

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Variable analysis

independent variables

Treatment with G. thunbergii

dependent variables

Expression of Bcl-2, Bax, caspase-3, and PARP genes

control variables

Cell lines: YCC-2 and SNU-668 cells
RNA extraction method: RNAiso Plus reagent (TaKaRa Bio, Japan)
CDNA synthesis method: cDNA Master Mix (ToYoBo, Japan)
Reverse transcription PCR protocol: as described previously (Kim et al. 2018)

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

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