Figure 4.

Exogenous FUS can rescue the effects of endogenous FUS depletion. (A) Western blot with FUS and GAPDH antibodies of protein samples from SK-N-BE cell lines carrying the construct FLAG-FUS^wt indicated in Figure 3B. These cells were treated with siRNA against the 3′UTR of FUS (siFUS-3′) for 6 days in RA and in the absence of Doxycycline. Scrambled siRNAs (siScr) were used as control. (B) Western blot of protein samples from SK-N-BE cell lines carrying the constructs indicated in Figure3B (FLAG-FUS^wt, FLAG-FUS^R521C and FLAG-FUS^P525L), together with a construct with no cDNA insertion (empty) treated as in (A) in the presence of Doxycycline. Exogenous FLAG-FUS expression was tested using Flag antibodies while GAPDH antibodies were utilized as control. (C) The histogram show the miRNA levels from cells treated as described in (A) and (B), analysed by RT–qPCR. Error bars represent s.e.m. from three independent measurements and the significance was assessed by Unpaired Student’s t-test (*P<0.05, **P<0.01, ***P<0.001). Source data for this figure is available on the online supplementary information page.