Fig. 2

LCL161 activates the non-canonical NF-κB pathway to enhance target gene transcription. a H1299 and A549 cells were treated with 10 μM/25 μM LCL161 or DMSO for 24 h. Cells were stimulated with 10 ng/ml TNFα for 5 min as a positive control. Phosphorylation of IκBα and IκBα was analysed by Western blotting. b After cells were treated with LCL161 or DMSO for 24 h, the expression levels of TRAF3, NIK, p100, p52, and cIAP1 were analysed by Western blotting. β-Actin served as the loading control. c NF-kB transcriptional activity was assessed by a luciferase assay, and the fold increase in luciferase activity is shown. d The expression of p100, p52 and RelB in both cytoplasm and nuclear were detected by Western blotting. Lamin B and tubulin served as the loading control. e The mRNA levels of some NF-κB target genes were examined by quantitative RT-PCR. Representative results are shown. Plots are the mean ± SEM of data from three independent experiments. *P < 0.05