Validation of genes identified through expression microarray profiling of 2D and 3D cultured FTSECs. The most significantly changing genes were validated by TaqMan qPCR in independent 2D and 3D cultures of FTSEC03, as well as in two additional cell lines from different patients (FTSEC01 and FTSEC284). Genes downregulated in 3D cultured cells (a) MARCH4 and (b) DIAPH3 were significantly downregulated in all 3 cell lines tested with the exception. (c) A similar trend was observed for GINS4 although this did not reach statistical significance. Genes upregulated in 3D cultured cells (d) C11orf96, (e) OLFM2A and (f) LRRK2 were all upregulated in all 3 samples following a transition to a 3D microenvironment. * P > 0.05, ** P > 0.01, *** P > 0.001, two-tailed paired Student’s T-tests were used to compare 2D and 3D cultured cells for each cell line individually.