Fig. 6

CircZBTB46 blocks the ubiquitination and degradation of hnRNPA2B1. (A) Western blot analysis of the expression level of hnRNPA2B1 after circZBTB46 knockdown or circZBTB46 overexpression. (B) RT‒PCR analysis of hnRNPA2B1 expression after transfection with si-circZBTB46 or the circZBTB46 overexpression plasmid. (C) Immunoblot analysis of the hnRNPA2B1 level in intact cells treated with CHX for 0, 3, 6 or 9 h. The relative fold changes were determined by comparison with the level at 0 h. (D) Cells transfected with si-circZBTB46 were treated with MG132 or CQ, and the hnRNPA2B1 level in the indicated cells was measured by Western blotting. (E) Lysates from circZBTB46-knockdown cells were immunoprecipitated (IP) with an anti-hnRNPA2B1 antibody and were then used for immunoblot analysis of ubiquitin and hnRNPA2B1. (F) Secondary structure prediction of circZBTB46 using the online web server RNAfold. The predicted binding sites of circZBTB46 with hnRNPA2B1 are shown in the blue circle. (G) The ubiquitination sites in hnRNPA2B1 were predicted via a Bayesian discriminant method and visualized using Discovery Studio 2021 (client version). Purple indicates the binding region, while yellow indicates the ubiquitination site