Fig. 6: Detection of cell cycle changes and treatment-related CalR translocation. | Cell Death Discovery

Fig. 6: Detection of cell cycle changes and treatment-related CalR translocation.

From: CDK inhibitors promote neuroblastoma cell differentiation and increase sensitivity to retinoic acid—a promising combination strategy for therapeutic intervention

Fig. 6

A, B Two-step cell cycle analysis: Cells were counted, adjusted to equal numbers, lysed, and stained with DAPI prior to measurement. DNA content was measured using the NucleoCounter® NC-3000™ system. Mean + SD; n = 3 biological replicates. One-way ANOVA (Tukey’s multiple comparisons test). *p < 0.05; ***p < 0.001; ****p < 0.0001 vs. ctrl. ##p < 0.01; ###p < 0.001; ####p < 0.0001 vs. monotherapy. C Apoptosis/necrosis assay. Flow cytometry was done to quantify the number of dead cells after 2 × 72 h treatment. Early apoptotic cells: Yo-Pro-1+/PI-, late apoptotic cells: Yo-Pro-1+/PI+, necrotic cells: Yo-Pro-1-/PI-. n = 3 biological replicates. One-way ANOVA (Tukey’s multiple comparisons test). *p < 0.05; **p < 0.01; ***p < 0.001 vs. ctrl; #p < 0.05; ##p < 0.01; ###p < 0.001 vs. monotherapy. D, E Immunogenic cell death analysis of NB cells (LAN-1, CHLA-90) was done by flow cytometry using PE anti-calreticulin antibody. D Representative flow cytometry plots of LAN-1 and CHLA-90 cells. E Quantitative analysis. A–E Doses: RA (both cell lines): 1.5 µM; LAN-1 abemaciclib: 0.2 µM; fadraciclib: 0.4 µM; CHLA-90: abemaciclib: 1.0 µM: fadraciclib: 1 µM; Mean + SD; n = 3 biological replicates. One-way ANOVA (Tukey’s multiple comparisons test). *p < 0.05; ***p < 0.001; ****p < 0.0001 vs. ctrl; #p < 0.05; ##p < 0.01; ###p < 0.001; ####p < 0.0001 vs. monotherapy.

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