Experimental validation of the effect of critical TFs and potential drugs in the hypoxia microenvironment. (A) Effects on tumor cell invasion ability after interfered CEBPD and FOSL1 with siRNA were examined by 3D sphere invasion assay in U87 and HG9 GBM cells under hypoxic conditions. Scale bar is 20 μm. Student's t-test, * P < 0.05, ** P < 0.01, *** P < 0.001. (B) The effect on tumor cell migration ability after interfered the CEBPD and FOSL1 with siRNA was examined by wound healing assay in LN229 GBM cells under hypoxic conditions. Scale bar is 50 μm. Student's t-test, * P < 0.05, ** P < 0.01, *** P < 0.001. (C) Effects on tumor cell invasion ability by Transwell invasion assay in U87, LN229 and HG9 GBM cells after interference of CEBPD and FOSL1 with siRNA under hypoxic conditions. Scale bars are 200 μm. Student's t-test, * P < 0.05, ** P < 0.01, *** P < 0.001. (D) Representative pictures and statistical plots of intracranial tumor size in mice after group treatment were shown by luciferase live imaging in an in situ tumorigenic mouse model constructed by U87 GBM cells, as well as statistics and analysis of survival in mice. Student's t-test and Log-rank (Mantel-Cox) test, * P < 0.05, ** P < 0.01, *** P < 0.001.