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NPAS3-HEK293-circadian-Illumina

ABSTRACT: The fragments of FLNPAS3 and DeltaNPAS3 were transferred into a similar TET-inducible expression plasmid and transfected into HEK293 [T-REx-293] cells (Invitrogen). At the zero hour time-point, cells were shifted into a serum-rich medium plus tetracycline in order to induce circadian cycling and NPAS3 overexpression. Parental HEK293 [T-REx-293] cells after circadian rhythmcity induction for +12hr/+24hrs were used as negative controls because stably integrated FLNPAS3/DeltaNPAS3 cell lines showed a low level of leaky transcription in the absence of tetracycline. At +2 hours, cells were washed with DMEM and then incubated with the same plus tetracycline for the remaining period of the experiment. Cell samples were collected at +12hrs and +24hrs respectively. For all circadian cell culture experiments with FLNPAS3/DeltaNPAS3/parental negative control cell lines, duplicate biological samples were assessed. Microarray probes synthesised from RNA extraction products was quantified using an Agilent Bioanalyzer to ensure high quality probes of equal quantity between experiments. Sentrix® HumanRef-8 v2 chips were used to detect gene expression profiles.

ORGANISM(S): Homo sapiens

SUBMITTER: Li Sha

PROVIDER: E-TABM-1079 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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