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. 2021 May 14;12(5):493. doi: 10.1038/s41419-021-03773-x

Fig. 5. miR-135b directly targets the deubiquitinase CYLD to activate NF-κB signaling.

Fig. 5

A Predicted miR-135b target sequence in CYLD-3′ UTR (WT) and mutant containing 4 mutated nucleotides in the seed sequence of CYLD-3′ UTR (CYLD-3′UTR-Mut). B Western blotting analysis of CYLD expression in the indicated cells. β-Tubulin served as a loading control. C, D The activity of the CYLD-3′ UTR-WT and CYLD-3′ UTR-Mut luciferase reporter gene in indicated cells. E Activity of the NF-κB luciferase reporter gene in the indicated cells transfected with NC, miR-135 mimics and miR-135 mimics plus CYLD gene (without 3′ UTR) or CYLD-3′ UTR. F Western blotting analysis of p-p65 and CYLD in the indicated cells transfected with vector, CYLD, or CYLD-3′ UTR. β-Tubulin served as a loading control. G Western blotting analysis of K63-linked poly-Ub of RIP1 in H292 and A549 cells.