Fig. 2

HMGCS2 and AKR1C3 expression is increased in PCa cells by incubation with CAF-conditioned medium. a mRNA expression of HMGCS2 and AKR1C3 was analyzed in LNCaP and DuCaP cells after 8 days of 3D spheroid culture in CAF-conditioned (CM) medium and compared to cells, which were cultured in standard medium. HMBS was used as internal control. Data represent the mean plus SEM from at least three independent experiments. (* P < 0.05, ** P < 0.01,*** P < 0.001) (b) Conditioned medium of CAF 3D spheroids was loaded onto a semi-quantitative RayBio® Human Cytokine Antibody Array (G-Series 1000, RayBiotech, Norcross, GA). Values were normalized to culture medium and expressed as mean signal intensity with SEM from three independent experiments. c CAFs were cultured either in T75 flasks (2D) or in 96 well hanging drop plates at 8000 cells per well. Alterations in mRNA gene expression were determined via Illumina microarray analysis. Significantly altered pathways between 2D and 3D cultured CAFs were identified via KEGG analysis. Representative phase contrast images are shown for CAFs grown either in 75cm² culture flasks (2D) or as 3D spheroids in 96 well hanging drop plates (magnification 10x, scale bar: 500 μm)