Fig. 6: Inhibition of NFATc1 sensitizes BLBC to conventional chemotherapy. | Cell Death & Disease

Fig. 6: Inhibition of NFATc1 sensitizes BLBC to conventional chemotherapy.

From: Decreased PRC2 activity supports the survival of basal-like breast cancer cells to cytotoxic treatments

Fig. 6

A qRT-PCR validation of NFATc1 knockdown. B Impairment of pG-2 cell growth upon NFATc1 knockdown determined by Celigo® (left panel) and crystal violet staining (right panel). C and D Reduction of pG-2 cells migratory capacity upon NFATc1 knockdown analyzed via gap closure assay (C) and Boyden chamber assay (D). E and F Assessment of EMT markers regulation in pG-2 cells upon NFATc1 knockdown by qRT-PCR. G The inhibition of NFATc1 activity by treating pG-2 cells with increasing concentrations of cyclosporin A (16, 80, 400 nM, and 2 µM) for 48 h increases the fraction of EpCAM positive cells, as measured by FACS. H Activation of the NFAT signaling through thapsigargin treatment (1, 5, 25, 125, and 625 nM) for 48 h reduces the fraction EpCAM-positive cells in pG-2 cells, as measured by FACS. I and J Inhibition of NFATc1 transcriptional activity by Cyclosporin A (CsA) (I) or VIVIT (J) treatment renders pG-2 cells more sensitive to CAF chemotherapy in vitro. Growth kinetics were assessed by Celigo® confluence measurement (left panel) and crystal violet staining (right panel). Error bars depicted as shadow area: standard error of the mean (SEM). K CAM assay demonstrating a significantly decreased size of the tumors treated with a combination of CAF + CsA. The Micro-CT scans of the tumors and quantification of their volumes are provided for the different conditions in the left and right panels, respectively. L–N Assessment of the CAF + CsA combination therapy efficiency in in vivo syngeneic mice transplanted with pG-2 cells. Graph depicting the treatment schedule for the different groups of mice (L). Growth kinetics of untreated (n = 3 animals) or CsA (50 mg/kg i.p. three times per week, n = 5 animals) treated tumors (M). Growth kinetics of tumors treated with either CAF alone (n = 3 animals) or CAF + CsA in combination (n = 3 animals) (N). *p < 0.05, **p < 0.001, ***p < 0.005. Student’s t-test: A, B, N (AUC: area under the curve), C–F, I, J, N. One-way ANOVA: G, H, K. All experiments were performed in at least biological triplicates (n = 3).

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