Fig. 6

The inhibitor of MYLK4 shows synergistic effect with the EGFR inhibitor in OS in vitro and in vivo. a 143B and b HOS cells treated with Gefitinib alone, ML-7 alone, or Gefitinib/ML-7 in combination. Cell viability was determined by CCK-8 assay. Combination index (CI) was calculated by using CompuSyn software. c Divide cells into four groups, Control group, Gefitinib alone group, ML-7 alone group and Gefitinib/ML-7 in combination group. Colony-formation assay was carried out in those four groups. Representative images of colony formation are shown in the upper panel. The numbers of cells in the indicated groups were shown in the histogram. The bars indicate the mean ± s.d. Statistically significant differences (t-test), **P < 0.01. d Wound healing assay was carried out in those four groups. Representative images of migration were shown in the upper panel. The degrees to which the wounds healed was shown in the histogram. The bars indicate the mean ± s.d. Statistically significant differences (t-test), **P < 0.01, Scale bars = 250 μm. e Transwell assay was carried out in those four groups. Representative images of invasion were shown in the upper panel. The proportions of invading cells were shown in the histogram. The bars indicate the mean ± s.d. Statistically significant differences (t-test), **P < 0.01, Scale bars = 200 μm. f The number of lung metastasis nodules formed by 143B cells in orthotopic osteosarcoma implanted mice was shown in the right panel, the mice were divided into four groups: mice treated with dmso only, mice treated with Gefitinib only, mice treated with ML-7 only and mice treated with Gefitinib plus ML-7. Statistically significant differences (t-test), **P < 0.01. Representative images of lung morphology and lung metastatic nodules in the four groups were shown in the left panel. Scale bars = 250 μm. g Tumor weights of the tumors in the four groups were recorded. Statistically significant differences (t-test), **P < 0.05. h The expressions of p-EGFR, p-AKT and p-ERK in vitro were measured by western blotting in different groups. i The expressions of p-EGFR, p-AKT and p-ERK in tumor xenograft tissues were measured by western blotting