Figure 2: Effects of FUS overexpression on miRNA levels.
From: An ALS-associated mutation in the FUS 3′-UTR disrupts a microRNA–FUS regulatory circuitry
(a,b) Levels of mature and pri-miRNA species for miR-141 and miR-200a measured by quantitative reverse transcriptase (qRT)-PCR in SK-N-BE cells (in absence or presence of Dox) stably transfected with either a RFP control (Ctrl) or RFP–FUS cDNAs. miR-15a and miR-432 as well as their corresponding pri-miRNA molecules were used as controls. Mature miRNA levels were normalized against snoRNA-U25 while pri-miRNAs against pre-GAPDH. (c) Left panel: representative western blot analysis of Zeb1 and FUS expression in SK-N-BE cells expressing Dox-inducible flag-FUS cDNA a. GAPDH was used as loading control. Right panel: densitometric analysis of Zeb1 normalized against GAPDH. See full blots with marker position in Supplementary Fig. 2c. (d) Lower panel: representative western blot analysis of FUS–WT and FUS–G48A expression in SK-N-BE cells transfected with the corresponding plasmids. Upper panel: densitometric analysis of FUS–WT and FUS–G48A normalized against GAPDH. See full blots with marker position in Supplementary Fig. 3a. (e) Levels of miR-141 and miR-200a measured by qRT-PCR in SK-N-BE cells treated as in panel d. miR-15a and miR-432 were used as controls. miRNAs levels were normalized against snoRNA-U44. All data were derived from three independent experiments; error bars represent s.e.m. *P<0.05 corresponds to the one-tailed Student’s t-test, **P<0.01, ***P<0.001.
