Fig. 4
From: NKX6-1 mediates cancer stem-like properties and regulates sonic hedgehog signaling in leiomyosarcoma
NKX6-1 promotes cancer stemness properties in LMS cells. Cells were seeded in ultralow attachment plates to assess sphere-forming ability. a Representative spheroid images from different NKX6-1 transfectants. NKX6-1-overexpressing MES-SA cells formed spheroids with round morphology (upper), and NKX6-1 knockdown MES-SA/DX5 cells formed clusters of loosely associated cells (lower). The scale bar represents 100 μm. b The number of spheres was quantified using ImageJ software. The total number of spheres increased upon NKX6-1 overexpression in MES-SA cells and decreased upon NKX6-1 knockdown in MES-SA/DX5 cells. The expression of cancer stemness-related genes correlated with sphere formation ability. c qRT-PCR was performed to evaluate stemness markers (including NESTIN, NANOG, KLF4, KLF8, MYC, OCT, SOX2, CD44, and CD49F) in MES-SA cells transfected with either NKX6-1 or vector control. d Expression of KLF8, MYC, and CD49F in MES-SA/DX5 cells transfected with either shNKX6-1 or scrambled control was determined by qRT-PCR. *p < 0.05