title: GUSB and ATP2B4 are suitable reference genes for CFTR gene expression data normalization in nasal epithelium cells.
abstract: BACKGROUND
  CFTR expression studies contribute in understanding the relationship between CFTR transcripts and clinical outcomes. Normalization of qPCR data is an essential step to determine target gene expression. Consequently, appropriate reference genes must be selected for each gene/tissue. In this work, we have assessed the suitability of four potential reference genes for CFTR expression analysis in nasal epithelium.
  
  
  METHODS
  B2M, GUSB, HPRT1 and ATP2B4 expression was evaluated in nasal epithelium samples (CFTR-wt controls, n=21; CFTR-splicing group, n=18) by RT-qPCR. Calibration curves were built and different analyses (geNorm, NormFinder, Mann-Whitney) were performed to evaluate gene expression stability between samples as well as between groups.
  
  
  CONCLUSIONS
  We have applied an accurate approach to select reference genes for CFTR expression analysis in nasal epithelium. From the four genes assessed, GUSB and ATP2B4 have been validated as a reliable gene combination for CFTR gene qPCR data normalization.
authors.affiliation: Human Molecular Genetics Group, IDIBELL, 08908 Barcelona, Spain.
authors.forename: Laia
authors.initials: L
authors.lastname: Masvidal
authors.forename: Antoni
authors.initials: A
authors.lastname: Alvarez
authors.forename: Laura
authors.initials: L
authors.lastname: Ruano
authors.forename: Javier
authors.initials: J
authors.lastname: de Gracia
authors.forename: Sara
authors.initials: S
authors.lastname: Larriba
authors.forename: Teresa
authors.initials: T
authors.lastname: Casals
mesh_terms: D000328:Adult; D003550:Cystic Fibrosis / Q000235:genetics / Q000473:pathology; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000235:genetics*; D020869:Gene Expression Profiling / Q000379:methods / Q000592:standards; D005966:Glucuronidase / Q000235:genetics*; D006801:Humans; D017136:Ion Transport / Q000235:genetics; D009297:Nasal Mucosa / Q000378:metabolism / Q000473:pathology; D053499:Plasma Membrane Calcium-Transporting ATPases / Q000235:genetics*; D012016:Reference Values; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D001613:beta 2-Microglobulin / Q000235:genetics*
chemical_list: D001613:beta 2-Microglobulin; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D005966:Glucuronidase; C507517:ATP2B4 protein, human; D053499:Plasma Membrane Calcium-Transporting ATPases