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. 2011 Jan 6;7(1):e1001269. doi: 10.1371/journal.pgen.1001269

Figure 5. hnRNP A1 co-regulates Mag alternative splicing.

Figure 5

(A) RNA was prepared from cells that had been transiently transfected with Hnrnpa1 or QK siRNA and selected with G418. RNA produced from CG-4 cells transfected with a control selection plasmid was analyzed alongside as a control. Isoforms of Mag containing or lacking exon 12 were amplified by RT-PCR using fluorescent primers. Products were separated in 5% acrylamide gels. Gels were scanned and the relative isoform ratio was calculated from the intensity profile of the lane using ImageGauge. Representative intensity profiles are presented below representative gel images. Data and standard deviations were calculated from 3 (control siRNA and Qk siRNA) or 5 (GFP control and Hnrnpa1 siRNA) independent transfections. An increase in expression of the longer isoform was detected upon transfection of both Hnrnpa1 and Qk siRNA. (B) Plp1 and Dm20 isoform ratios were analyzed as described above for Mag. A decease in expression of the upper isoform was detected when Qk siRNA was transfected but not when Hnrnpa1 siRNA was transfected. (C) Qk and Hnrnpa1 knockdown were confirmed by qRT-PCR.