Fig. 2: PC3-CAFs organoid growth: effect of Rh-2025u peptide.

AR-negative PC-derived PC3 cells stably expressing GFP were cultured in ECM, in the presence of pooled AR-positive CAFs from different patients (Table 2), for 3 days (basal condition). Cells were then left unchallenged or challenged for 15 days 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 a, the organoid size was monitored by phase-contrast (left section), or fluorescence microscopy (middle section). Merged images are shown in the right section. Scale bar, 100 μm. In b, the organoid size was calculated as described in the legend to Fig. 1 and expressed as fold increase in the relative PC3-CAFs organoid size. In c and d, GFP-PC3 cells were cultured in ECM, in the absence of CAFs. In c, the organoid size was monitored on 3 and 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 d, GFP-PC3 cells in ECM were cultured for 3 days (basal condition). The 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. The organoid size was calculated as described in the legend to Fig. 1 and expressed as fold induction in the relative PC3 organoid size. In b and d, n represents the number of experiments. Means and SEM are shown. *p < 0,05, as compared to basal or untreated cells.