Fig. 4

c-Jun enhances miR-17-92 promoter transcription activity. A miR-17-92 promoter region (positions 5786–8494 in accession# NG-032702) was inserted into pEZX-LvPG04 dual-reporter vector (WT promoter). This vector uses GLuc (Gaussia Luciferase) as the promoter reporter and SEAP (secreted alkaline phosphatase) as the internal control for signal normalization (top panel). The potential two c-Jun binding sites in the promoter were deleted (MT promoter) (middle panel). Negative control uses empty vector (Empty promoter) (bottom panel). B Relative luciferase (Luc) activity in the culture medium from HEK293T cells transfected with the miR-19b promoter vector (promt) or empty negative vector (empty) together with transfection with c-Jun-expression vector (c-Jun-ov) or control vector (ctrl). Mean ± SE for three experiments. C Relative luciferase activity in the culture medium from HEK293T cells transfected with the miR-19b promoter vector (promt) or empty negative vector (empty) then exposed to H/R or normoxia (Norm). Mean ± SE for four experiments. D Western blot analysis of c-Jun and GAPDH proteins. E Relative luciferase (Luc) activity in the culture medium from HEK293T cells transfected with the miR-19b promoter vector (promt) or empty negative vector (empty) together with transfection with c-Jun-siRNA (cJun-si) or scrambled RNA (scRNA). Mean ± SE for three experiments. F Relative luciferase (Luc) activity in the culture medium from HEK293T cells transfected with wild-type miR-19b promoter vector (wild) or mutated vector (mut) together with transfection with c-Jun-expression vector (cJun-ov) or control vector (ctrl). Mean ± SE for three experiments