MRNA expression profiles of SK-N-AS neuroblastoma cells with induced ALK-wt, ALK-F1174L or ALK-R1275Q

ABSTRACT: Establishment of inducible SK-N-AS ALKwt, ALKF1174L and ALKR1275Q cell lines Human neuroblastoma SK-N-AS cells were electroporated with pcDNA6/TR (Invitrogen) using the NeonM-eM-( Transfection System (Life Techologies). Single cell clones were generated using blasticidin (7.5M-eM-5g/ml) and limited dilution. Using a TetR antibody (Clonetech), the clone with the highest TetR expression was selected (named SK-N-AS-TR) and used for the transfection with pT-REx-DEST30-ALK, ALKF1174L or ALKR1275Q. After transfection of SK-N-AS-TR with the ALK variants, single cell clones were raised using geneticin (500 M-eM-5g/ml) and limited dilution, while blasticidin treatment was continued as described above. Total RNA of (treated) cell lines, transgenic mice tumors and ganglia was isolated using the miRNeasy kit (Qiagen) according to the manufacturerM-^IM-[M-*s instructions, including on-column DNase treatment. Samples were labeled and hybridised to the Sureprint G3 human GE or G3 Mouse GE 8x60K microarrays (Agilent Technologies), according to the manufacturerM-^IM-[M-*s guidelines and starting from 200 ng of RNA.

ORGANISM(S): homo sapiens

SUBMITTER: Katleen De Preter 

PROVIDER: E-MTAB-3207 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress