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. 2021 Jan 12;12:295. doi: 10.1038/s41467-020-20527-z

Fig. 4. circNDUFB2 acts as a scaffold to enhance the binding of IGF2BP proteins with TRIM25.

Fig. 4

a Binding of circNDUFB2 with TRIM25 or HECTD3 in A549 cells. b RNA FISH-immunofluorescence showed the co-localization of circNDUFB2 (red) with TRIM25 (green) in A549 cells. Scale bar = 50 μm. c Co-immunoprecipitation (Co-IP) showed the binding of TRIM25 with IGF2BPs in A549 cells. d Immunofluorescence detected the co-localization of IGF2BPs (red) with TRIM25 (green) in A549 cells. Scale bar = 50 μm. e Protein levels of IGF2BPs in A549 cells with TRIM25 knockdown. f Protein levels of IGF2BPs in A549 cells with TRIM25 or TRIM25-mutant (TRIM25ΔRBD) overexpression. g After TRIM25 overexpression, immunoblot detected protein levels of IGF2BPs in A549 cells with or without circNDUFB2 knockdown. h–j Co-IP showed the binding of TRIM25 with IGF2BP proteins. Two independent experiments were carried out with similar results. RNase A (10 μg/ml), RNase R (100 U/ml).