NCM460-NTR1 cells were generated from human colonic epithelial NCM460 cells (INCELL, San Antonio, TX, USA) transduced with lentivirus particles as previously described 10 (link) and maintained in M3:D culture media (INCELL) supplemented with 10% fetal bovine serum (FBS, Life Technologies, Grand Island, NY, USA). Human embryonic kidney fibroblasts, HEK293, and colonic cancer HCT-116 cells were maintained in Eagle’s Minimum Essential Medium (MEM, Life Technologies) and McCoy5a (ATCC, Manassas, VA, USA) respectively and supplemented with 10% FBS. Lipofectamine 2000, lipofectamine RNAimax and OptiMEM were from Life Technologies. Neurotensin was from Bachem Americas (Torrance, CA, USA). TNBS and SR48962 were from Sigma Aldrich (St. Louis, MO, USA) and DSS was from MP Biomedicals (Santa Ana, CA, USA). Goat anti-AFTPH (sc-167055), mouse anti-villin and rabbit anti-β tubulin were from Santa Cruz Biotechnology (Santa Cruz, CA, USA).
Protocol detail
Modeling Colonic Epithelial Cell Lines
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NCM460
M3:D culture media
MEM
McCoy5a
Lipofectamine 2000
lipofectamine RNAimax
OptiMEM
Neurotensin
SR48962
DSS
rabbit anti-β tubulin
Cells
Colonic
Colonic cancer
Eagle
Embryonic
Epithelial cells
Fibroblasts
Goat
Hct 116 cells
Human
Kidney
Lentivirus
Lipofectamine
Lipofectamine 2000
Mouse
Neurotensin
Rabbit
Tubulin
Villin
Corresponding Organization :
Other organizations : University of California, Los Angeles
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Variable analysis
independent variables
- Transduction of NCM460 cells with lentivirus particles
- Generation of NCM460-NTR1 cells
- Cell maintenance and growth in M3:D culture media
- Human embryonic kidney fibroblasts (HEK293) maintained in Eagle's Minimum Essential Medium (MEM) with 10% FBS
- Colonic cancer HCT-116 cells maintained in McCoy5a with 10% FBS
- Lipofectamine 2000, lipofectamine RNAimax and OptiMEM from Life Technologies
- Neurotensin from Bachem Americas
- TNBS and SR48962 from Sigma Aldrich
- DSS from MP Biomedicals
- Goat anti-AFTPH (sc-167055), mouse anti-villin and rabbit anti-β tubulin from Santa Cruz Biotechnology
- Not specified
- Not specified
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