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. 2013 Jul 4;8(7):e69096. doi: 10.1371/journal.pone.0069096

Figure 2. AON-induced ALK2 exon 8 skipping in various cell types.

Figure 2

(A) C2C12 cells were transfected with 500 nM non-targeting, fluorescently-labeled AONs fluorescent AON in differentiation medium and fluorescent images were taken 2 days after transfection. (B) Various of cells were transfected with indicated control AON or ALK2 AON. RNA was isolated 2 days after transfection, and RT-PCR was performed to visualize the full length Alk2 (composed of exon 7, 8 and 9); and skipped Alk2 (composed of exon 7 and exon 9). (C) MEECs cells were transfected with 100 nM control AON or 100 nM ALK2 AON, RNA was isolated 1 day after transfection. C2C12 cells and KS483 cells were transfected with AONs in proliferation medium; RNA was isolated 2 days post transfection. cDNA was synthesized using random hexamer primers and used for the quantification of Alk2 expression. The relative full length ALK2 mRNA expression was analyzed. Alk2 RNA level was normalized to Gapdh; the level of expression for untreated sample was defined as 1. Values and error bars represent the means ± SD of triplicates. Statistical analysis was performed using Student’s t-test. *P<0.05, **P<0.005.