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

Figure 8.

Figure 8

circCdyl acts as a sponge for let-7c and promotes C/EBP-δ expression in macrophages

(A) Results of luciferase reporter assays. RAW264.7 cells were cotransfected with let-7c mimic or mimic-NC and a luciferase reporter containing wild-type circCdyl (circCdyl-WT) or mutant circCdyl (circCdyl-mut). Luciferase activity was analyzed after 24 h. ∗∗p < 0.01; n = 3 per group. (B) Results of luciferase reporter assays. RAW264.7 cells were cotransfected with let-7c mimic or mimic-NC and a luciferase reporter containing wild-type C/EBP-δ (C/EBP-δ-WT) or mutant C/EBP-δ (C/EBP-δ-mut). Luciferase activity was analyzed after 24 h. ∗∗p < 0.01; n = 3 per group. (C) Results of luciferase reporter assays. RAW264.7 cells were cotransfected with or without let-7c mimic, oe-circCdyl or oe-vector, and C/EBP-δ-WT or C/EBP-δ-mut. Luciferase activity was analyzed after 24 h. ∗∗p < 0.01; n = 3 per group. (D and E) C/EBP-δ protein levels in RAW264.7 cells transfected with mimic-NC, let-7c mimic, inhibitor-NC, or let-7c inhibitor and then stimulated with LPS+IFN-γ (24 h). ∗∗p < 0.01; n = 5 per group. (F and G) C/EBP-δ protein levels in RAW264.7 cells transfected with oe-circCdyl or oe-vector and then stimulated with LPS+IFN-γ (24 h). ∗∗p < 0.01; n = 5 per group. (H and I) C/EBP-δ protein levels in RAW264.7 cells transfected with oe-vector + mimic-NC, oe-circCdyl + mimic-NC, oe-vector + let-7c mimic, or oe-circCdyl+let-7c mimic and then stimulated with LPS + IFN-γ (24 h). ∗∗p < 0.01; n = 5 per group. (J and K) C/EBP-δ protein levels in RAW264.7 cells transfected with si-scramble + inhibitor-NC, si-circCdyl + inhibitor-NC, si-scramble+let-7c inhibitor, or si-circCdyl+let-7c inhibitor and then stimulated with LPS+IFN-γ (24 h). ∗∗p < 0.01; n = 5 per group.