DLL4 is targeted by circHIPK3 and miR-30c(A) qPCR analysis of DLL4 levels in TREx cells 0 and 24 hours post induction.Figure 5(A) qPCR analysis of DLL4 levels in TREx cells 0 and 24 hours post induction. GAPDH was used a housekeeper (n=3). Data information: data are presented as mean ± SD. *P<0.05, **P<0.01, ***P<0.001 (Unpaired Student's t-test). All repeats are biological. (B) Representative western blot of DLL4 expression in TREx cells 0 and 24 hours post induction. GAPDH was used a loading control (n=3). Data information: data are presented as mean ± SD. *P<0.05, **P<0.01, ***P<0.001 (Unpaired Student's t-test). All repeats are biological. (C) qPCR of DLL4 levels in scr and circHIPK3 KD stable TREx cells at 0 and 24 hours post induction with GAPDH as a housekeeper (n=3). Data information: data are presented as mean ± SD. *P<0.05, **P<0.01, ***P<0.001 (Unpaired Student's t-test). All repeats are biological. (D) Representative western blot of DLL4 levels in scr and circHIPK3 stable TREx cells at 0 and 24 hours post induction with GAPDH as a loading control. Densitometry analysis of n=3. Data information: data are presented as mean ± SD. *P<0.05, **P<0.01, ***P<0.001 (Unpaired Student's t-test). All repeats are biological. (E) qPCR analysis of DLL4 levels in TREx cells that have been transfected with either scr or miR-30c 24 hours pre-induction, analysis performed 0 and 24 hours post induction. GAPDH was used a housekeeper (n=3). Data information: data are presented as mean ± SD. *P<0.05, **P<0.01, ***P<0.001 (Unpaired Student's t-test). All repeats are biological. (F) Representative western blot of DLL4 levels in scr or miR-30c transfected TREx cells at 0, 24 and 48 hours post induction with GAPDH as a loading control. Densitometry analysis of n=3 Data information: data are presented as mean ± SD. *P<0.05, **P<0.01, ***P<0.001 (Unpaired Student's t-test). All repeats are biological. (G) Fold enrichment of GAPDH and DLL4 in biotinylated miR-30c RIPs over Scr transfected into HEK-293Ts. (n=3) Data information: data are presented as mean ± SD. *P<0.05, **P<0.01, ***P<0.001 (Unpaired Student's t-test). All repeats are biological. (H) Luciferase Reporter Assay from HEK-293Ts co-transfected with DLL4 3'UTR reporter plasmid and either scr or miR-30c. Data presented is relative to an internal firefly control (n=3). Data information: data are presented as mean ± SD. *P<0.05, **P<0.01, ***P<0.001 (Unpaired Student's t-test). All repeats are biological. (I) qPCR analysis of DLL4 levels in TREx cells that have transfected with either scr or a miR-30c antagomiR 24 hours pre-induction, with analysis performed at 0 and 24 hours post induction. GAPDH was used as a housekeeper (n=3). Data information: data are presented as mean ± SD. *P<0.05, **P<0.01, ***P<0.001 (Unpaired Student's t-test). All repeats are biological. (J) Representative western blot of DLL4 levels in scr or miR-30c antagomiR transfected TREx cells at 0, 24 and 48 hours post induction with GAPDH as a loading control. Densitometry analysis of n=3. Data information: data are presented as mean ± SD. *P<0.05, **P<0.01, ***P<0.001 (Unpaired Student's t-test). All repeats are biological. (K) qPCR analysis of DLL4 expression in Ago2 RIPs as a percentage of input. TREx Cells were transfected with either a scrambled control or a miR-30c antagomiR. (n=3) Data information: data are presented as mean ± SD. *P<0.05, **P<0.01, ***P<0.001 (Unpaired Student's t-test). All repeats are biological. (L) Luciferase Reporter Assay from HEK-293Ts co-transfected with DLL4 3'UTR reporter plasmid and either scr or miR-30c antagomiR. Data presented is relative to an internal firefly control (n=3). Data information: data are presented as mean ± SD. *P<0.05, **P<0.01, ***P<0.001 (Unpaired Student's t-test). All repeats are biological.