Fig. 9

Pharmacological and genetic inhibition of LDHA in MDA MB231 cells. a MDA PA and MDA RES cells were treated with GSK for 48 h. Relative cell viability was analyzed. b, c MDA PA and MDA RES cells were treated with 25 and 50 μM GSK for 48 h; ATP (b) and NAD(H) (c) levels were determined, respectively. d Synergistic effect of FK866 and GSK inhibits cell growth in MDA RES cells. MDA RES cells were treated with various concentrations of GSK and co-treatment with 20 nM FK866 for 48 h. Percentage of cell viability was determined. e, f Downregulation of LDHA by siRNA was performed in MDA RES cells. Twenty-four hours after silencing, MDA RES cells were treated with GSK for 48 h. Expression levels of ATF4 (e) and CHOP (f) were evaluated. Expression of GAPDH was used as a housekeeping gene. All data were represent as mean ± S.D. from three independent experiments (*p < 0.05, **p < 0.01, ***p < 0.001). g MDA PA and MDA RES cells were performed anchorage-independent growth (colony formation assay) in soft agar along with indicated concentrations of FK866 and GSK treatment for 21 days. A representative colony formation experiment was illustrated and colony number was counted by operetta