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. 2017 Mar 30;8:14741. doi: 10.1038/ncomms14741

Figure 2. CircRNA expression upon FUS depletion and in MN differentiation of mESCs.

Figure 2

(a) circRNA expression analysed by qRT–PCR in sorted GFP+-FUS+/+ and GFP+-FUS−/− cells. The 14 downregulated (left) and 5 upregulated (right) species are reported. CircRNA levels were normalized against Atp5o mRNA levels and expressed as relative quantity with respect to the GFP+-FUS+/+ sample set to a value of 1. (b) The histogram shows the expression level of circRNAs, measured by qRT–PCR, in FUS+/+ mESCs and GFP+-FUS+/+ and GFP−-FUS+/+ cells. Their values were normalized against Atp5o mRNA levels and expressed as relative quantity with respect to GFP− samples set to a value of 1. For a,b, error bars represent s.e.m. of three independent experiments. *P<0.05, **P<0.01 and ***P<0.001 correspond to two-tailed Student's t-test. (c) Left panel: location on the circRNA of the DIG-labelled probe used for northern blot analysis. Right panel: Northern blots on 10 μg of total RNA from FUS+/+ mESCs, GFP−-FUS+/+, GFP+-FUS+/+ and GFP+-FUS−/− cells. The circular forms are indicated aside the gels; asterisks indicate two additional bands likely corresponding to the two alternative linear forms of the c-31 host mRNAs (Hdgfrp3-001, ENSMUST00000107305.7, 5,887 nt; Hdgfrp3-002, ENSMUST00000026094.5, 2,865 nt). Each blot is shown in parallel to the EtBr staining of the agarose gel, where the migration of the 18S and 28S rRNAs is indicated.