NSC12 targets WM cells by inhibiting NF-κB activity. (A) BCWM.1 and MWCL.1 cells were exposed to NSC12 (6 μM) for 12 hours and subjected to wide transcriptome profiling, showing a significant inhibition of NF-κB signaling-related gene sets, as shown by performing GSEA (P < .05; FDR < 25%). (B) BCWM.1 and MWCL.1 cells were cultured with NSC12 (0 to 6 μM), for 4 hours, and then exposed TNF-α (10 ng/mL) was added for the last 20 minutes. NF-κBp65 transcription factor binding to its consensus sequence on the plate-bound oligonucleotide was studied from nuclear extracts. Wild-type and mutant are wild-type and mutated consensus competitor oligonucleotides, respectively. All results represent means (± SD) of triplicate experiments. (C) BCWM.1 and MWCL.1 cells were cultured in the presence or absence of NSC12 (0 to 6 μM), for 4 hours, and then exposed TNF-α (10 ng/mL) was added for the last 20 minutes. WM cells were then harvested, and cell lysates were subjected to western blot using anti-p-IkBa, -IkBa, and -actin antibodies. (D) BCWM.1 and MWCL.1 cells were cultured in the presence or absence of NSC12 (0 to 6 μM), for 4 hours, and then exposed TNF-α (10 ng/mL) was added for the last 20 minutes. WM cells were then harvested, and nuclear protein lysates were extracted and subjected to western blot using anti-p-p65, -p65, -RelB, -p100, -p52, -p50, -p105, and -nucleolin antibodies.