Fig. 4

HMGA1 increases the transcriptional activity of FOXM1 on VEGFA promoter. a Luciferase assay on HEK293T cells transiently co-transfected with the luciferase reporter plasmid pGL4.10-VEGFprom (− 1000–1) and with pEGFP-FOXM1 and increasing quantities of pEGFP-HMGA1 expression vectors (red bars). pRL-CMV Renilla luciferase expression vector was included to normalize for transfection efficiencies. Values are reported as relative luciferase activity compared to cells transfected with pEGFP-FOXM1 in the absence of pEGFP-HMGA1 and are presented as the mean ± SD (n = 3). **p < 0.01, ***p < 0.001; two-tailed Student’s t-test. Below the graph, the correspondent western blot validations of HMGA1 and FOXM1 protein levels are reported. b Luciferase assay on HEK293T cells silenced for HMGA1 by siRNA and transiently co-transfected with the luciferase reporter plasmid pGL4.10-VEGFprom (− 1000–1) with the expression plasmid pEGFP-FOXM1. pRL-CMV Renilla luciferase expression vector was included to normalize for transfection efficiencies. Values are reported as relative luciferase activity compared to cells transfected with the control expression vector pEGFP. The data are represented as the mean ± SD (n = 3), *p < 0.05; two-tailed Student’s t-test. Below the graph, the correspondent western blot validation. β-actin was used as a loading control for endogenous HMGA1