tissue: Colon cell line: HCT116 cell type: colon cancer cell treatment: LS-1-2 time: 24H after treatment
Treatment protocol
HCT116 cells treated with 0.1μM LS-1-2 was conducted at 6 h , 12 h, and 24 h after treatment
Growth protocol
Human CRC cell lines HCT116 were 158 purchased from American Type Culture Collection and maintained in Dulbecco’s 159 Modified Eagle medium (Gibco, Carlsbad, CA) with 10% fetal bovine serum, 100 U/mL 160 penicillin sodium, and 100 mg/mL streptomycin sulfate in humidified 5% CO2 at 37 °C.
Extracted molecule
total RNA
Extraction protocol
Total RNA from six cell samples was extracted using TRIZOL Reagent (Cat#15596-018, Life Technologies, Carlsbad, CA, US) following the manufacturer’s instructions and checked for a RIN number to inspect RNA integrity by an Agilent Bioanalyzer 2100 (Agilent Technologies, Santa Clara, CA, US). Qualified total RNA was further purified by RNeasy micro kit (Cat#74004, QIAGEN, GmBH, Germany) and RNase-Free DNase Set (Cat#79254, QIAGEN, GmBH, Germany).
Label
biotin
Label protocol
Total RNA was amplified, labeled, and purified by Affymetrix WT PLUS Reagent Kit (Cat#902280, Affymetrix, Santa Clara, CA, US) according to the manufacturer’s instructions to obtain biotin-labeled cDNA.
Hybridization protocol
The labeled cDNA targets were then hybridized on slides of Affymetrix GeneChip® Human Transcriptome Array 2.0.
Scan protocol
Slides were scanned Affymetrix GeneChip® Scanner 3000 (Cat#00-00213, Affymetrix, Santa Clara, CA, US).
Description
HCT116 cells treated with 0.1μM LS-1-2 was conducted at 24 h
Data processing
Data were extracted using Command Console Software 4.0 (Affymetrix, Santa Clara, CA, US).
