Fig. 5

TRIM59 was transcriptionally regulated by YAP1/TEAD4. A Immunofluorescence staining was performed to detect YAP1 or TEAD4 expression and distribution during hypoxia. Amplification: 400 ×. B The promoter sequence of TRIM59 was inserted into pGL3-basic plasmid. Then the luciferase reporter assay was performed to determine the effect of YAP1 and TEAD4 on the transcription of TRIM59. C Differently truncated fragments of TRIM59 promoter were inserted into pGL3-basic plasmid. Then the luciferase reporter assay was performed to determine the effect of YAP1 and TEAD4 on the transcription of TRIM59. D ChIP assay was conducted to verify the bind between TEAD4 and TRIM59 promoter. IgG served as the negative control. E The binding of TRIM59 promoter and TEAD4 was confirmed by DNA pull down assay. F In order to explore the effect of YAP1 and TEAD4 on the transcription of TRIM59, PASMCs were infected with adenovirus carrying YAP1 shRNA or TEAD4 shRNA. Then the levels of YAP1 and TEAD4 were measured by qPCR. G After infection, the mRNA level of TRIM59 was determined by qPCR. H Western blot was performed to detect the protein level of TRIM59. *p < 0.05