Fig. 6

CYLD is identified as a direct and functional target of miR-181b. a Western blot analysis of CYLD in SFCs and Eca109 parental cells treated with vector pcDNA3.1 STAT3 or siSTAT3. b CYLD expression level in SFCs quantified by qPCR. c Bioinformatics analysis conservation of CYLD 3′–UTR among different species and CYLD 3′-UTR is a target of miR-181b. d Diagram of CYLD 3′-UTR-containing reporter construct. Mutations were generated at the three predicted miR-181b binding sites located in the CYLD 3′-UTR. e Luciferase reporter assays of wild-type and mutant reporter plasmids cotransfected with miR-181b or NC into SFCs dissociated with trypsin. f qPCR analysis of CYLD in SFCs treated with miR-181b mimic or miR-181b inhibitor. g Western blot analysis of CYLD expression in SFCs treated with miR-181b mimic or miR-181b inhibitor. h NF-κB activity detection in SFCs treated with miR-181b mimic or miR-181b inhibitor. i IL-6 activity detection in SFCs treated with miR-181b mimic or miR-181b inhibitor. β-Actin was used as an internal control. Error bars represent mean ± SD