MUCs were cultured in DMEM/F12 GlutaMAX™ with 10% fetal bovine serum (FBS, all from Invitrogen) in an incubator supplied with 5% CO2 at 37°C as previously described [16 (link)]. When MUCs reached approximately 80–90% confluence, 5-aza-2′-deoxycytidine (5-aza-CdR, Sigma) was added to culture medium at 1, 2, and 4 μM. In the control group, vehicle (DEME/F12) was applied to MUC cultures. MUCs were observed daily using phase contrast microscopy and digital images were captured using a digital camera. After 48 hr of 5-aza-CdR treatment, half of the culture medium was replaced and MUCs were maintained for another 24 hr. In this study we investigated the effect of 5-aza-CdR on MUC gene expression at 72 hr after treatment, which was determined based on previous publications [30 (link),32 (link), 36 (link)].
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Epigenetic Modulation of MUC Gene Expression
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5 aza
5 aza 2 deoxycytidine
After treatment
Camera
Culture medium
Deme
Gene expression
Phase contrast microscopy
Corresponding Organization :
Other organizations : Wayne State University
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Variable analysis
independent variables
- Concentration of 5-aza-2'-deoxycytidine (5-aza-CdR) (1, 2, and 4 μM)
- MUC gene expression at 72 hr after treatment
- Culture medium (DMEM/F12 GlutaMAX™ with 10% fetal bovine serum (FBS))
- Incubator conditions (5% CO2 at 37°C)
- Vehicle (DMEM/F12) applied to control group
- Positive control: Not mentioned.
- Negative control: Vehicle (DMEM/F12) applied to control group.
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