Fig. 7

Characterization of MDA MB231 and FK866-resistant MDA MB231 cells. a, b MDA MB231 parental (MDA PA) and resistant cells (MDA RES) cells were treated with 10 and 100 nM FK866 for 48 h. ATP (a) and NAD(H) (b) levels were measured. c Western blot showing no alter in translation inhibition in MDA RES along with FK866 treatment in respect to MDA PA. Protein expression from total lysates were determined by detecting with indicated antibodies. B-ACTIN was used as a loading control. d MDA PA and MDA RES cells were treated with 20 nM FK866 for 48 h. Expression of KYNE and IDO, genes involved in the de novo pathway for NAD⁺ synthesis, were evaluated. e–h Relative cell viability of CEM PA and CEM RES cells treated with JPH203 (e), 2-deoxyglucose (f), L-Asp (g), and oligomycin A (h) for 48 h was displayed. i Analysis of mitochondria content. MDA PA and MDA RES cells were exposed to mitotracker staining. FCCB was used as a negative control. j Expression of genes involved in glycolysis and OXPHOS pathway were determined in MDA PA and MDA RES cells. Expression of 18S was served as a housekeeping gene. k Enhanced LDHA activity in the resistance. Enzymatic activity of glycolysis pathway was measured in MDA PA and MDA RES cells. Data were plotted as mean ± S.D. from three biological experiments with technical triplicates (*p < 0.05, **p < 0.01, ***p < 0.001)