Fig. 1: NPB inhibits BAD Serine (S) 99 phosphorylation and reduces cell survival.
A Cell viability assay: Dose-response curve of University of Mysore (UM) supplied NPB in the year 2020. The cell survival fraction (%) was measured at 72 h (n = 3). The IC50 value was calculated using GraphPad Prism (7.0). B Western blot: The pBADS99 and BAD protein levels in MCF-7 cells treated with NPB as indicated for 48 h and evaluated using WB analysis. C The pBADS99 and BAD protein levels in MCF-7 cells treated with the NPB dose as indicated for 24, 48, or 72 h and evaluated using WB analysis. V=vehicle. β-ACTIN was used as an input control. D The non-ionic detergent Tween-80 enhances the efficacy of NPB. Cell viability (%) was analyzed by using MTT and Alamar Blue assays. Data represent mean ± SD (n = 3). * (#)P < 0.05, ** (##)P < 0.01, and *** (###)P < 0.001 compared. P-value in the t-test compared to TWEEN-80 is denoted as #, P-value in the t-test compared to NPB (10 μM) with 0% TWEEN-80 is denoted as *. UM: University of Mysore. E Live/dead assay performed using 3D Matrigel culture assays previously performed1,2,3,5,10,11,12. The illustration was generated using the online platform BioRender.com. The unprocessed western blot images are represented in Supplementary Fig. 3.
