Fig. 4: RS-35d, S-35d and R-35d differential effect in PDAC 3D cultures. | Cell Death Discovery

Fig. 4: RS-35d, S-35d and R-35d differential effect in PDAC 3D cultures.

From: Investigating synthetic lethality and PARP inhibitor resistance in pancreatic cancer through enantiomer differential activity

Fig. 4

a Cell viability measured after 72 h exposure to 40 µM RS-35d, 20 µM S-35d or 20 µM R-35d alone or in combination with 10 µM olaparib (OLA) in BxPC-3 3D spheroids. b I. index of RS-35d, S-35d or R-35d association with olaparib in BxPC-3 3D spheroids. I. index values were made explicit when the drug association showed synergism (i. index < 0.8). c Evaluation of the effect of the 72 h treatment with 40 µM RS-35d alone or in combination with 10 µM OLA on BxPC-3 3D spheroid volume and cell death; time-course brightfield and fluorescence images of BxPC-3 3D spheroids treated as previously described (scale bar, 200 μm); time-course analysis of the corresponding 3D spheroid volume (expressed as % of the spheroid volume at t = 0 h); analysis of cell death in 3D spheroid at 72 h (expressed as PI/Calcein-AM ratio). Results are expressed as mean ± SD (n = 6). Evaluation of cell viability (d) and apoptosis (e) upon 120 h treatment with 40 µM RS-35d alone or in combination with 10 µM OLA in human PDAC organoids; time-course representative images of RS-35d- and RS-35d/olaparib-induced apoptosis via caspase 3/7 cleavage (scale bar, 400 μm) and analysis of the corresponding caspase 3/7 cleavage-induced green fluorescence signal. Results are expressed as mean ± SD (n = 3). Statistical analysis was performed with two-way ANOVA (a, c, d, e) or one-way ANOVA (c) followed by Tuckey’s multiple comparison test, with §p < 0.05, §§p < 0.01 or §§§p < 0.001 vs CTRL; ##p < 0.01 or ###p < 0.001 vs 10 µM OLA; *p < 0.05 or **p < 0.01 vs RAD51-BRCA2 inhibitor alone.

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