Fig. 6: Lnc408-mediated CBY1 decrease triggers WNT signaling and promotes self-renewal and stemness maintenance of BCSCs. | Cell Death & Disease

Fig. 6: Lnc408-mediated CBY1 decrease triggers WNT signaling and promotes self-renewal and stemness maintenance of BCSCs.

From: A novel Lnc408 maintains breast cancer stem cell stemness by recruiting SP3 to suppress CBY1 transcription and increasing nuclear β-catenin levels

Fig. 6

A A protein networks showing the potential interaction among 14-3-3 (YWHAZ), β-catenin (CTNNB1), and CBY1. B HEK293T cells were transfected with an expression vector encoding wild-type CBY1 (WT) or mutant CBY1 (S20A). Immunoprecipitation and western blotting were conducted with antibodies against 14-3-3, β-catenin, and CBY1 to detect the interaction of CBY1, 14-3-3, and β-catenin. C IP-WB was used to detect the ternary complex of CBY1, 14-3-3, and β-catenin in non-CSC derived from of BT549, non-CSC transfected with ectopic lnc408 (oelnc408), and non-CSC with silenced CBY1 (sh-CBY1); or in CSC derived from BT549, CSC transfected with ectopic CBY1 (oeCBY1), and CSC with silenced lnc408 (sh-lnc408). D Western blotting to check the cellular distribution of β-catenin as in C (C cytoplasm, N nucleus). GAPDH served as a cytoplasm control, and PCNA as a nucleus control. E The mammosphere-forming abilities of BT549, BT549 transfected with CBY1 (oeCBY1), or BT549/ocCBY1 cells treated with or without SLK2001 (WNT/β-catenin agonist, 10 μM) were tested under suspended culture. The down panel shows the statistical results of mammosphere numbers as means ± SD (**P < 0.01; NS none significance; scale bar, 100 μm). F, G The mRNA and protein expressions of breast cancer pluripotent factors (KLF4, c-Myc) were determined by qRT-PCR (F) and western blotting (G), respectively (*P < 0.05; **P < 0.01; NS none significance).

Back to article page