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. Author manuscript; available in PMC: 2021 Oct 9.
Published in final edited form as: Methods Mol Biol. 2021;2372:193–202. doi: 10.1007/978-1-0716-1697-0_17

Fig. 2.

Fig. 2

Validation of circRNA interacting proteins. (a) Biotinylated antisense oligomers (ASOs) complementary to the junction of circSamd4 and control (Ctrl) were incubated with C2C12 in GM as well as in DM. After affinity pulldown using streptavidin beads, the levels of circSamd4 enrichment in ASO pulldown samples were assessed by RT-qPCR analysis of circSamd4 (relative to the enrichment of GAPDH mRNA, a transcript that does not bind the ASOs and encodes a housekeeping protein) in the pulldown samples. Enrichment in Samd4 mRNA was monitored in parallel. (b) Top 5 proteins shared in the mass spec datasets from mouse and human myoblast pulldowns. (c) Validation in C2C12 of other proteins identified as being selectively enriched in circSamd4 and circSAMD4 ASO pulldowns. Following ASO pulldown, the presence of candidate proteins was assessed by Western blot analysis. Input, 5 μg. (Figure is reproduced from ref. 15)