Fig. 7.

Reduction of SMN decreases the relative amount of the mgU2-30 fragment from ectopically expressed scaRNA9. (A) Northern blot of RNA from HeLa cells transfected with control (lane 2), SMN-A (lane 3) or SMN-B (lane 4) siRNA for 24 h and then transfected with pcDNA 3.1+ scaRNA9 for 24 h. A histogram was created quantifying the ratio between the processed mgU2-30 fragment and ectopic full-length from six different biological repeats. (B) Northern blot of RNA from HeLa cells transfected with SMN-B siRNA for 24 h followed by the co-transfection of scaRNA9 and GFP plasmid (lane 1) or scaRNA9 and GFP-SMN plasmid (lane 2) for 24 h. A histogram was created quantifying the ratio between mgU2-30 and ectopic full-length from eight different biological repeats. (C) Western blot showing SMN knockdown and rescue. HeLa cells were transfected with negative control or SMN-B siRNA for 24 h and then co-transfected with GFP plasmid (lanes 1 and 2) or with GFP-SMN plasmid (lanes 3 and 4) for 24 h. The membrane was then probed for GFP and SMN, respectively. β-tubulin was then probed as a loading control. Standard error was used to generate error bars. Student’s t-test was used to determine statistical significance, indicated by * corresponding to a P-value<0.05 (B); *** corresponding to a P-value<0.0005 (A).