Expression of CTCF in leukemic cells before and after treatment with an activator of NF-κB activity. (A) Nalm-6 cells were treated with different doses (5 or 10 μg/ml) of the NF-κB activity activator LPS for 12 h. NF-κB pathway activity was detected by Western blot using a nuclear p65-specific antibody. Histone H3 and α-tubulin were used as the loading controls for nuclear and cytoplasmic proteins, respectively. (B) CTCF mRNA levels in cell lysates were measured by real-time PCR. The relative changes in the expression levels after treatment with 5 and 10 μg/ml LPS were analyzed and presented as mean ± SD from triple replications (fold change 1.33, p = 0.027; fold change 1.66, p = 0.025, respectively). (C) Cell lysates were probed with an anti-CTCF antibody, and the expression levels of CTCF were semi-quantified by analyzing the Western blot with Gel-Pro Analyzer software. The fold changes in expression after treatment with 5 and 10 μg/ml LPS were 1.53 and 1.72, respectively. GAPDH was used as a loading control.