Fig. 1: LNCaP-CAFs organoid growth: effect of Rh-2025u peptide.

LNCaP cells stably expressing GFP were used. In a, cells were analyzed by phase contrast (left section) and fluorescence microscopy (middle section). Merged images are shown in the right section. Scale bar, 10 μm. AR-positive CAFs from different patients (Table 2) were pooled and used. In b and c, GFP-LNCaP cells in ECM were co-cultured with CAFs for 3 days (basal condition). Cells were then left unchallenged or challenged with 10 nM R1881, in the absence or presence of 10 nM Rh-2025u peptide (Rh), or 10 μM enzalutamide (enz). Control cells were treated with the peptide or enzalutamide, alone. In b, the organoid size was monitored for 15 days and representative images by phase-contrast (left section), or fluorescence microscopy (middle section) were captured and shown. The corresponding merged images are shown in the right section. Scale bar, 100 μm. In c, the organoid size was calculated using the Leica Suite software under basal conditions (3 days) or in cells left unstimulated or stimulated for 15 days, in the absence or presence of the indicated compounds. It was finally expressed as a fold increase in the relative LNCaP-CAFs organoid size. n represents the number of experiments. Means and SEM are shown. *p < 0,01, as compared to basal or untreated cells.