96 well plates

Cells (8000/well) were seeded into 96-well plates (Servicebio, WuHan, China), and proliferation was evaluated using Cell Counting Kit-8 (Biosharp Biotechnology, Beijing, China) reagent. The optical density was read at 450 nm. For clone formation assays, cells were cultured for 14 days, fixed with 4% paraformaldehyde for 15 min, and stained with 0.1% crystal violet for 15 min (Meilunbio, Dalian, China). For scratch assays a linear wound was created in a monolayer of serum-starved cells using a 10-μL pipette tip, and cell coverage across this line was determined. For Transwell migration assays, cells were seeded into the upper chamber (8 μm; BIOFIL, Guangzhou, China) and incubated with serum-free medium. The lower chamber contained medium with 10% FBS. After 24 hours, cells were fixed with 4% paraformaldehyde and stained with 0.1% crystal violet.

We mainly used a ridge regression-based method of the R package oncoPredict (version 0.2) to predict the IC50 AUC values of potential drug responses which bridges the in vitro drug screening with in vivo drug and biomarker discovery [17 (link)]. The Oxaliplatin sensitivity of the transfected CRC cells was detected using CCK-8 assays. First, cells that were seeded in 96-well plates (Servicebio, Wuhan, China) were incubated with 10 µM Oxaliplatin (Selleck, China) for 24 h. Next, 10 µl of CCK-8 reagent (Dojindo, Kumamoto, Japan) was added, and the cells were incubated at 37 °C for 2 h. Finally, the absorbance was measured at 450 nm in an ELISA 96-well microtiter plate reader (Bio-Rad 680, California, USA).