A colony formation assay was performed as described previously [47 (link), 52 (link)]. Approximately equal amounts of PANC-1 cells transfected with vector, wild-type or mutant TRPM8 were seeded in 12-well plates and allowed to grow for 7~10 days. The medium was replaced every 3 days. Cells were washed twice with PBS, fixed with 4% paraformaldehyde, and stained with 0.5% crystal violet staining solution (Sigma-Aldrich, USA). Colonies with more than 50 cells in triplicate wells were counted.
Huang Y., Li S., Liu Q., Wang Z., Li S., Liu L., Zhao W., Wang K., Zhang R., Wang L., Wang M., William Ali D., Michalak M., Chen X.Z., Zhou C, & Tang J. (2022). The LCK-14-3-3ΞΆ-TRPM8 axis regulates TRPM8 function/assembly and promotes pancreatic cancer malignancy. Cell Death & Disease, 13(6), 524.
Other organizations :
Hubei University of Technology, Tongji Hospital, Huazhong University of Science and Technology, Zhengzhou Children's Hospital, Zhengzhou University, Wuhan University, Renmin Hospital of Wuhan University, University of Alberta
Genetic manipulation of PANC-1 cells: vector, wild-type TRPM8, or mutant TRPM8
dependent variables
Colony formation and growth of PANC-1 cells
control variables
Seeded equal amounts of PANC-1 cells in 12-well plates
Allowed cells to grow for 7-10 days
Replaced medium every 3 days
Washed cells twice with PBS
Fixed cells with 4% paraformaldehyde
Stained cells with 0.5% crystal violet
controls
Positive control: wild-type TRPM8
Negative control: vector
Annotations
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