Journal: Oncogene

Article Title: Identification of CASZ1 nuclear export signal (NES) reveals potential mechanisms for loss of CASZ1 tumor suppressor activity in neuroblastoma

doi: 10.1038/onc.2016.179

Figure Lengend Snippet: (A) IPA assay of CASZ1b targets in SY5Y cells determined by microarray showed gene enrichment in the categories of molecular and cellular functions with a key subcategory being cell growth and proliferation (threshold represents p=0.05). (B) GSEA assay indicated the positive enrichment of genes involved neurological system process (left panel), and negative enrichment of genes regulated by MYC (right panel). NES, normalized enrichment score; Nom, nominal; FDR, false discovery rate. (C) Transcriptional activity assay of cytoplasmic localized K37A mutant and loss of NuRD binding mutant L31A in SY5Y cells by realtime PCR. a, Left panel: western blot analysis of CASZ1b and mutant proteins from SY5YtetCASZ1b and CASZ1b mutant cells after 24 h Tet treatment; right panel: realtime PCR analysis of CASZ1b and mutant mRNA levels from SY5YtetCASZ1b and CASZ1b mutant cells after 24 h Tet treatment. b-f, realtime PCR result verified the regulation of TH, NGFR, CD9, MYC and KIT by CASZ1b in SY5Y cells after 24 h Tet treatment. Compared to wild type CASZ1b, K37 almost completely lost transcriptional activity at regulating all these genes in SY5Y cells (data are shown as means ± S.E.M., *: p<0.05). Compared to wild type CASZ1b, L37 significantly decreased transcriptional activity at regulating TH, NGFR and MYC, but had a slight increase at regulating CD9 and no effect on KIT (data are shown as means ± S.E.M., *: p<0.05).

Article Snippet: Gene set enrich analysis (GSEA) ( , ) was performed using the whole gene list generated by uploading the raw microarray data to Partek Genomics Suite 6.6.

Techniques: Microarray, Activity Assay, Mutagenesis, Binding Assay, Western Blot