Figure 4.

LC3B promotes NANOG-dependent aggressive phenotypes through the AKT signaling pathway. (A-D) CaSki-no insert cells and CaSki-NANOG cells were transfected with siGFP, siLC3B #1 or siLC3B #2 then incubated in medium supplemented with 0.1% FBS for 48 h. (A) The levels of LC3B, p-AKT1, AKT1, CCNA1, and MCL1 protein in these cells were analyzed by western blots. (B-C) Flow cytometry analysis of the frequency of apoptotic (active CASP3⁺) cells (B) after incubation with or without MLANA-specific CTLs at a 1:1 ratio for 4 h or (C) after intracellular delivery of GZMB for 4 h. (D) The degree of CSC-like phenotypes was assessed by the sphere-forming capacity in low-density suspension culture. All experiments were performed in triplicate. *p < 0.01 and **p < 0.001, by two-way ANOVA (B, C and D). The data represent the mean ± SD