Fig. 5

ANGPT2 is required for HOXD9-mediated proliferation, cell cycle progress and apoptotic inhibition of NSCLC cells. NCI-H292 cells were co-transfected with siHOXD9 and the plasmid expressing ANGPT2 (exANGPT2) or empty vector. NCI-H661 cells were co-transfected with exHOXD9 and siANGPT2 or siNC. A After 48 h of transfection, ANGPT2 mRNA expression in two NSCLC cells (NCI-H661 and NCI-H292) was evaluated by real time-PCR assay. B After 48 h of transfection, ANGPT2 protein expression in two NSCLC cells (NCI-H661 and NCI-H292) was evaluated by western blot assay. C After 48 h of transfection, NSCLC cells were seeded into 96-well microplates and analyzed with CCK-8 reagents at indicated time. D Cell cycle progression in two NSCLC cells was evaluated by flow cytometry analysis. E Apoptosis in NCI-H292 cells was evaluated by flow cytometry analysis. The apoptotic rate was quantified by adding the percentages of early apoptotic cells (Annexin V⁺/PI⁻ in Q4 quadrant) and late apoptotic cells (Annexin V⁺/PI⁺ in Q2 quadrant). * p < 0.05; ** p < 0.01; *** p < 0.001. Data in (A), (C), (D) and (E) are presented as mean values ± SD. The blots were cropped and the original uncropped images of blots were shown in supplementary materials