Ex z0396 m02

Human cDNA CD133 expression plasmid (EX-Z0396-M02) and empty vector plasmid (EX-NEG-M02) were obtained from GeneCopoeia. Lentiviral shRNA pGIPZ vectors; NS (RHS4348) and αCD133 (V2LHS_71816) were obtained from Thermo Scientific. IKK (IKK-2 S177E S191E) and IKBα (pBabe-Puro-IKBalpha-mut (superrepressor)) plasmids were obtained from Addgene.

Authenticated human embryonic kidney (HEK) 293 cells and human colon cancer cell lines (LoVo and LS174T cells) were purchased from ATCC, immediately prepared frozen cell stocks and stored in liquid nitrogen freezer. Cells were passaged for fewer than 6 months after resuscitation. Cell line authentication was performed by ATCC using short tandem repeat DNA profiles. All cell lines were routinely maintained in ATCC-recommended conditions. 293-CD133 cells overexpressing CD133 were established by transfection with CD133-expressing plasmid, pcDNA3.1-CD133 (CD133 cDNA cloned into pcDNA3.1, Invitrogen), and were maintained with G418 (600 μg/ml, Invitrogen). Cells were incubated in a 37°C and 5% CO2 environment under humidified conditions. Human cDNA CD133 expression plasmid (EX-Z0396-M02) and empty vector plasmid (EX-NEG-M02) were obtained from GeneCopoeia.

Human cDNA CD133 expression plasmid (EX-Z0396-M02) and empty vector plasmid (EX-NEG-M02) were obtained from GeneCopoeia. Lentiviral shRNA pGIPZ vectors; NS (RHS4348) and aCD133 (V2LHS_71816) were obtained from Thermo Scientific.
MIA PaCa-2 (ATCC) and stable MIA-derivatives were maintained in DMEM (Hyclone) containing 10% fetal bovine serum. S2-VP10 cells were cultured in RPMI 1640 (Hyclone) supplemented with 10% fetal bovine serum. Stable clones were selected and maintained in Geneticin (Invitrogen) and Puromycin (Clontech) for MIA PaCa-2 and S2-VP10 derivatives, respectively.