Fig. 4.

Effect of GANT61 on proliferation, apoptosis and hedgehog pathway. Inhibitory effect of GANT61 on proliferation and apoptosis of breast cancer cells was tested using in vitro models. Cell viability assays were conducted using CCK-8, to observe the effect of GANT61 on cell proliferation in MDA-MB-231 and MCF-7 in a. Dose dependent (5, 10, 15, 20 μM) and b. Time dependent manner (24, 48, 72, 96 h). Apoptosis was assessed using Annexin V at variable concentrations (5, 10, 15, 20 μM) of GANT61 in c.MDA-MB-231 and d.MCF-7 (Unpaired t test, *p < 0.05, **p < 0.001, ***p < 0.0001). IC50 of GANT61 was determined to be 10 μM. Inhibition of expression of hedgehog pathway after treatment with GANT61 (10 μM) for 48 h. Relative mRNA expression of SHH, PTCH1 and GLI1 using qPCR in e. MDA-MB-231, f. MCF-7 (Mann Whitney U test, *p < 0.05, **p < 0.001, ***p < 0.0001). g. Protein expression of SHH, PTCH1 and GLI1 using western blot having β-actin as internal control in both cell lines. All experiments were conducted in triplicates, performed thrice and values are represented as mean ± S.D.