Fig. 2

ATF5 upregulation in bladder cancer cells enhances a TIC-like phenotype. A GSEA of correlations between ATF5 gene expression and stemness-related gene signatures (YAMASHITA_LIVER_CANCER_STEM_CELL_UP, LEE_NEURAL_CREST_STEM_CELL_UP, JAATINEN_HEMATOPOIETIC_STEM_CELL_DN, RAMALHO_STEMNESS_DN). B Western blotting detection of ATF5 in the indicated ATF5-upregulation, ATF5-knockdown, or control cells. GAPDH was used as the loading control. C, D qRT-PCR (C) and WB (D) detection of stemness-related markers (ABCG2, OCT4, SOX2) in overexpressing, downregulating ATF5 or control cells. E, F Representative images and quantitative analyses of tumor sphere formation in ATF5-upregulation cells (E, × 100), ATF5-knockdown cells (F, × 100) or control cells. G Tumors formed by ATF5-overexpressing bladder cancer cells were larger than control tumors. In contrast, tumors formed by ATF5-knockdown cells were smaller than those formed by RNAi-vector cells. Representative images of tumors were displayed (left). Tumor growth curves after injection of indicated cells (right). Bars represent the mean ± SD of three independent experiments; *P < 0.05, **P < 0.01, ***P < 0.001. GSEA, gene set enrichment analysis; TIC, tumor initiating cells; qRT-PCR, quantitative real-time polymerase chain reaction; WB, western blotting