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. 2020 Jan 25;48(7):3789–3805. doi: 10.1093/nar/gkaa035

Figure 2.

Figure 2.

Characterization of circSamd4. (A) Schematic of exon 3 of Samd4 mRNA, located on mouse chromosome 14 and giving rise to a 519-nucleotide long circSamd4, and detail of the sequence junction. (B) RT-qPCR results showing the abundance of circSamd4 and Samd4 mRNA in total RNA from C2C12 cells (GM) that were either left untreated or treated with RNase R. RNA levels were normalized to the levels of Gapdh mRNA. (C) Relative levels of Samd4 mRNAs as measured by RT-qPCR analysis in GM and DM (72 h) C2C12 cultures. RNA levels were normalized to the levels of Gapdh mRNA in the same samples. (D) Relative levels of circSamd4 in the nuclear (Nuc) and cytoplasmic/sarcoplasmic (Cyto) compartments of C2C12 GM and DM (72 h) cultures; the quality of the fractionation was assessed by monitoring the levels of a predominantly cytoplasmic transcript (Actb mRNA) and a predominantly nuclear lncRNA (7sk). (E) Brightfield micrographs to visualize circSamd4 (red) and Samd4 mRNA (blue) in proliferating (GM) and differentiated (DM) C2C12 cultures using the BaseScope single-molecule detection system (Advanced Cell Diagnostics, CA) (Materials and Methods). Nuclei are counterstained with hematoxylin. Data in (B, C) are the means ±SEM from three or more independent experiments. **P < 0.005.