Journal: Molecular Therapy

Article Title: Long Non-coding RNA BGas Regulates the Cystic Fibrosis Transmembrane Conductance Regulator

doi: 10.1038/mt.2016.112

Figure Lengend Snippet: BGas and as4 mediated regulation of CFTR . ( a ) A schematic depicting the CFTR locus with transcriptional start sites (TSS) for CFTR , EST BG213071 (BGas), and those primers used to evaluate CFTR expression. The sasRNA target site (as4) in the BGas promoter is also shown along with the two exons making up BGas. ( b ) The effects of BGas over-expression using an exogenously expressed BGas (exBGas) on CFTR expression in 1HAEo- cells. The control (pcDNA3.1) and exBGas transfected cells are shown. ( c,d ) The effects of sasRNA as4 relative to control (pU6M2) on ( c ) BGas and ( d ) CFTR expression in 1HAEo- cells. ( e,f ) The effects of exBGas ( e ) and sasRNA as4 ( f ) on CFTR expression in CFPAC cells. ( g ) Dose-dependent effect of an siRNA targeting the as4 site (siRNA4) on ΔF508- CFTR chloride channel transport function in CF Human Bronchial Epithelial (CFhBE) primary cells (epithelial voltage clamp assay, Ussing chamber). ( h ) The localization of biotin containing BGas at the intergenic locus of CFTR in transfected CFPAC cells contrasted with the lambda biotin control. For b–f , the averages of triplicate treated cultures are shown with the standard error of the means and a P value from a paired T -test, * P < 0.05 and ** P < 0.01.

Article Snippet: Briefly, primary human bronchial epithelial (hBE) cells from a CF patient homozygous for ΔF508- CFTR (Asterand Biosciences, Royston, UK) were plated onto Costar Snapwell (Corning, Corning, NY) tissue-culture inserts coated with collagen type IV, and cultured in Bronchial epithelial growth medium (Lonza, Basel, Switzerland).

Techniques: Expressing, Over Expression, Transfection

Journal: Molecular Therapy

Article Title: Long Non-coding RNA BGas Regulates the Cystic Fibrosis Transmembrane Conductance Regulator

doi: 10.1038/mt.2016.112

Figure Lengend Snippet: BGas and as4 mediated regulation of CFTR . ( a ) A schematic depicting the CFTR locus with transcriptional start sites (TSS) for CFTR , EST BG213071 (BGas), and those primers used to evaluate CFTR expression. The sasRNA target site (as4) in the BGas promoter is also shown along with the two exons making up BGas. ( b ) The effects of BGas over-expression using an exogenously expressed BGas (exBGas) on CFTR expression in 1HAEo- cells. The control (pcDNA3.1) and exBGas transfected cells are shown. ( c,d ) The effects of sasRNA as4 relative to control (pU6M2) on ( c ) BGas and ( d ) CFTR expression in 1HAEo- cells. ( e,f ) The effects of exBGas ( e ) and sasRNA as4 ( f ) on CFTR expression in CFPAC cells. ( g ) Dose-dependent effect of an siRNA targeting the as4 site (siRNA4) on ΔF508- CFTR chloride channel transport function in CF Human Bronchial Epithelial (CFhBE) primary cells (epithelial voltage clamp assay, Ussing chamber). ( h ) The localization of biotin containing BGas at the intergenic locus of CFTR in transfected CFPAC cells contrasted with the lambda biotin control. For b–f , the averages of triplicate treated cultures are shown with the standard error of the means and a P value from a paired T -test, * P < 0.05 and ** P < 0.01.

Article Snippet: Briefly, primary human bronchial epithelial (hBE) cells from a CF patient homozygous for ΔF508- CFTR (Asterand Biosciences, Royston, UK) were plated onto Costar Snapwell (Corning, Corning, NY) tissue-culture inserts coated with collagen type IV, and cultured in Bronchial epithelial growth medium (Lonza, Basel, Switzerland).

Techniques: Expressing, Over Expression, Control, Transfection