Metabolical and physiological analyses comparing 2D with 3D culture. [A] Energy metabolism: lactate accumulation measurements in 3D compared to 2D culture. Data are expressed as ratio μmoles lactate/ μg protein. [B] Western immunoblotting showing HIF1a protein expression both in 2D and 3D cultures at day 4 and 7. [C] mRNA expression of target genes downstream HIF1a in 2D and 3D cultures. Real Time-PCR data are calculated as expression ratio 3D/2D. A representative experiment out of three is shown. [D] Western blotting shows Glut1 protein expression both in 2D and 3D cultures on day 4 and 7. GAPDH is used as loading control.