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. 2021 Sep 20;30(2):915–931. doi: 10.1016/j.ymthe.2021.09.017

Figure 4.

Figure 4

circCdyl binds to IRF4 and inhibits its entry into the nucleus

(A) Silver-stained sodium dodecyl sulfate polyacrylamide gel electrophoresis gel of proteins immunoprecipitated by circCdyl and its antisense circRNA. The red box indicates the region of the gel that was excised and processed for mass spectrometry. IRF4 protein expression was assayed by western blotting. (B) RNA immunoprecipitation (RIP) experiments were performed using an antibody against IRF4 or negative immunoglobulin G (IgG). ∗∗p < 0.01 versus IgG; n = 3 per group. RIP-derived RNA was measured by qPCR analysis, and IRF4 was expressed as a percentage of the input. (C) Total IRF4 mRNA expression in peritoneal macrophages in which circCdyl was inhibited or overexpressed (detected by qPCR). NS, not significant; n = 5 per group. (D and E) Total IRF4 protein levels in peritoneal macrophages in which circCdyl was inhibited or overexpressed (detected by western blotting, β-actin as the internal reference). NS, not significant; n = 5 per group. (F and G) Cytoplasmic and nuclear IRF4 protein levels in peritoneal macrophages stimulated with IL-4 and overexpressing circCdyl (detected by western blotting, β-actin as the internal reference). ∗∗p < 0.01; n = 5 per group. (H and I) Cytoplasmic and nuclear IRF4 protein levels in peritoneal macrophages stimulated with IL-4 and silenced for circCdyl (detected by western blotting, β-actin as the internal reference). ∗∗p < 0.01; n = 5 per group.