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. 2022 Nov 7;2(11):1355–1371. doi: 10.1158/2767-9764.CRC-22-0245

FIGURE 6.

FIGURE 6

Characterization of immortalized human prostate cancer cell line ACRJ-PC28 growing as 3D organoids. A, ACRJ-PC28 cells were seeded on and embedded in Matrigel as described in the Materials and Methods and monitored over time by phase contrast microscopy. Organoids are shown at days 7 and 10. Lens 10×. Scale bar: 400 μm. Both ACRJ-PC28 and hPrEC-C1 cells form round organoids, but ACRJ-PC28 show lumens. PC-3 organoids form invasive protrusions (protrusions are marked with white arrows on day 7, and a green arrow marks the same organoid with many more protrusions). B and C, IF staining with CK5, CK18, and p63, and AR antibodies and imaged by confocal microscopy, 63×, demonstrates expression of basal markers CK5 and p63. However, expression of luminal marker CK18, and AR was negative. PC-3 and LNCaP organoids were used as positive controls for CK18 and AR, respectively. D, ACRJ-PC28 organoid vimentin and E-cadherin expression patterns suggest an opposite graded relative expression (designated H:high, I:intermediate, L:low). E, Expression of laminin β1 and integrin β1. B–E, Primary antibody incubation was followed by staining with fluorescence conjugated secondary antibody and phalloidin. Lens: 63×. Scale bar: 100 μm. Zoom factor: 0.75.