Figure 2. NFκB is responsible for maintaining CSC characters in suspension- and hypoxia-cultured cells.

A. High levels of NFκB p65 and HIF2α protein were detected in the nuclear protein extracted from suspension- and hypoxia-cultured cells. Nuc: nucleolin used as a loading standard. B. Phosphorylated AKT, NFκB p65_Ser276 and degradation of IκBα were detected in suspension- and hypoxia-cultured cells by western blot. α-Tub: α-tubulin used as a loading control. C. High NFκB DNA binding activity was detected by EMSA. Mut and WT: mutant and wild type probe competition. D and E. High NFκB p65 protein (D) and transcriptional activity (E) were detected in p65 transfected clones (C1, C3, P1 and C2) by western blot and luciferase reporter gene assay respectively. Mock: empty vector transfected cells. F. High ALDH⁺ and CD24^low/CD44^high population was detected in NFκB p65 transfected clones.