Fig. 2.

Comparison of content and localization of relevant proteins within the various morphological structures formed by the P69, M12, F6 and M12-Vim prostate sublines grown embedded in lrECM gels. (A) A Z-stack of P69 acini from day 8 3D cultures was examined by immunofluorescence confocal microscopy. These 8 pictures represent plane 4, 8, 12, 16, 20, 24, 28 and 32 from 40 such planes. Acini were fixed and stained with antibody to α6-integrin (green). All pictures are taken at a magnification of 63X and nuclei were counterstained with DAPI (blue) as discussed in Materials and Methods. (B) Confocal immunofluorescence microscopy of structures formed by the various sublines at day 8 in lrECM stained with antibodies to vimentin (green) and Ki-67 (red), panels a₁ to a₄; β-catenin (red), panels b₁ to b₄; and E-cadherin (green), panels c₁ to c₄; keratin 5/6 (green), panels d₁ to d₄; keratin 8 (green), panels e₁ to e₄; and nuclei (blue) in all panels. (C) Cellular structures were stained with antibodies to α6-integrin (green), panels h₁ to h₄, or β1-integrin (red), panels j₁ to j₄ with overlay of α6β4 integrin in panel k₄. For details of immunostaining see Materials and Methods. (D) LnCap and PC3 cell lines were grown in lrECM and stained at day 8 as described in panel C. For panels B, C and D a size marker of 5 µm is shown.