Journal: eLife
Article Title: EPB41L4A-AS1 long noncoding RNA acts in both cis - and trans -acting transcriptional regulation and controls nucleolar biology
doi: 10.7554/eLife.106846
Figure Lengend Snippet: RT-qPCR to assess the expression of the reported genes after ( A ) transfection with siRNAs against EPB41L4A-AS1, ( B ) CRISPRa with guides targeting the EPB41L4A-AS1 promoter, ( C ) transfection with plasmid encoding the EPB41L4A-AS1 cDNA, transfection with GapmeRs ( D ) and siRNAs ( E ) targeting EPB41L4A, ( F ) CRISPRa with guides targeting the EPB41L4A promoter, and ( G ) transfection with plasmid encoding the EPB41L4A cDNA. ( H ) Changes in EPB41L4A-AS1 expression after rescuing EPB41L4A-AS1 with an ectopic plasmid or CRISPRa following its KD with GapmeRs. In both panels (Ectopic OE and CRISPRa) the ‘-’ samples represent those transfected with the Empty Vector or sgControl. Asterisks indicate significance relative to the –/– control (transfected with both the control GapmeR and vector). ( I ) Same as in ( H ), but for changes in EPB41L4A expression. ( J ) UMI-4C contact profiles in control and LNA2-transfected cells using baits targeting the TSS of EPB41L4A-AS1. The green area represents the quantified genomic interval, and the p-value was calculated using a Chi-squared test. All experiments were performed in n = 3 biological replicates, except UMI-4C with n = 2, with the error bars in the boxplots representing the standard deviation. In all cases, ns = p > 0.05; *p < 0.05; **p < 0.01; ***p < 0.001 (two-sided Student’s t -test).
Article Snippet: RNA was then retrotranscribed using the qScript Flex cDNA Synthesis Kit (Quanta Bio), following standard conditions as suggested by the manufacturer.
Techniques: Quantitative RT-PCR, Expressing, Transfection, Plasmid Preparation, Control, Standard Deviation