Dataset: Transcription profiling of human LNCaP cell lines transfected with AIbZIP
Androgen-induced bZIP (AIbZIP) is a basic leucine zipper (bZIP); transcription factor that is encoded by the CREB3L4 (cAMP responsive...
Androgen-induced bZIP (AIbZIP) is a basic leucine zipper (bZIP); transcription factor that is encoded by the CREB3L4 (cAMP responsive element; binding protein 3-like 4) gene. In humans, AIbZIP mRNA is most abundant in; the prostate but it is also present, albeit at much lower levels, in the; pancreas and other tissues. The full-length AIbZIP protein localizes to the; endoplasmic reticulum and the available data suggest that AIbZIP is; processed to its transcriptionally active form by regulated intramembrane; proteolysis (RIP). To define the transcriptional programme that is initiated; by AIbZIP, we generated LNCaP prostate cancer cell lines that conditionally; express the processed form of AIbZIP. The recombinant protein consists of; the first 290 amino acids of AIbZIP fused to a C-terminal HA epitope. Conditional expression was achieved using the RheoSwitch expression system. Clone number 7-11 was exposed to the inducing agent (RSL1) for 6 and 24 hr; and Affymetrix microarrays were used to identify AIbZIP-regulated; transcripts. Candidate genes were identified by comparing the probe set; intensities of RSL1-treated cells and control cells (cells exposed to; vehicle). Forty-eight genes were then validated by Northern blot analysis. Experiment Overall Design: Clone 7-11 cells (passage 42) were plated in 24 10-cm culture dishes. Experiment Overall Design: Sixteen dishes were cultured in medium containing RSL1 (500 nM) and eight Experiment Overall Design: dishes were cultured in medium containing an equivalent amount of the RSL1 Experiment Overall Design: vehicle (DMSO). Eight RSL1-treated dishes were harvested 6 hr after addition Experiment Overall Design: of RSL1 and another eight dishes were harvested 24 hr after addition of Experiment Overall Design: RSL1. At each time point, the dishes were pooled two by two to yield a total Experiment Overall Design: of 12 individual samples i.e. 4 controls, 4 exposed to RSL1 for 6 hr, and 4 Experiment Overall Design: exposed to RSL1 for 24 hr. One sample of each group was processed to nuclear Experiment Overall Design: extracts to assess the levels of recombinant AIbZIP by immunoblotting. The Experiment Overall Design: three other samples of each group were processed separately to cRNA probes Experiment Overall Design: which were then hybridized to Affymetrix Human Genome U133 Plus 2.0 arrays Experiment Overall Design: (Batch nos. 4003850 and 4003847).
- Dec.12, 2014
- Sep.21, 2014