Fig. 2: Transcriptionally active NF-κB is increased in the nucleus of TKI-resistant CML cells.

A Bar graphs represent relative luciferase reporter activity in K562^S versus K562^R cells (n = 5 samples/group) stably transduced with an NF-κB reporter versus a negative control. B Immunoblot shows NF-κB and pNF-κB in the nucleus and cytoplasm of K562^S versus K562^R cells. C Bar graph represents densitometric calculations for separate replicates (n = 3) of the nucleocytoplasmic fractionation experiments presented in panel (B). Densitometric analysis was completed by normalizing against lamin B and tubulin in the nuclear and cytoplasmic fractions, respectively. D Immunoblot shows NF-κB and pNF-κB in the nucleus and cytoplasm of CD34⁺ cells from newly diagnosed, TKI-sensitive CP-CML patients (n = 2) versus TKI-resistant patients (n = 3) and normal donors (n = 2). Lamin B and tubulin were used to assess the quality of separation of the nuclear and cytoplasmic fractions, respectively. E Bar graph represents densitometric calculations for pNF-κB in panel (D). Error bars represent standard error of mean (SEM). C cytoplasmic fraction, N nuclear fraction, T total fraction.