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. 2016 Jul 6;7:84. doi: 10.3389/fendo.2016.00084

Figure 2.

Figure 2

Expression of carbonic anhydrase 9 (CA9) in 2D and 3D cultures of PC3 cells grown alone or in transwell coculture with adipocytes. Immunofluorescence analysis of CA9 expression (green) in monolayer PC3 cultures grown alone (A) or in transwell with adipocytes (B). (C) 2D no primary CA9 antibody control; DAPI was used as nuclear marker (Blue). 3D reconstruction of CA9 expression in tumor spheroids grown alone (D) or in transwell with adipocytes (E); green arrow (X), red arrow (Y), and blue arrow (Z) indicate orientation of the spheroid in 3D space. Middle slice through z-stack showing CA9 expression in spheroids cultured alone (F,G,J,K) or in transwell with adipocytes (H,I,L,M). Hoechst dye was used for labeling nuclei [(F,H); blue], CA9 [(G,I); green], DIC [(J,L); differential interference contrast], and merged images (K,M). (N) 3D culture; no CA9 primary antibody control; 40× images; bar, 50 μm.