Effects of lycorine on phosphorylation of IκB and nuclear import of NF-κB. (A) Effect of lycorine on expression of IκB and phosphorylation of IκB protein. (B) Effect of lycorine on expression of NF-κB protein and (C) distribution of NF-κB protein in nucleus. α-tubulin was used for normalisation and verification of the protein loading in A, B, and C in Western blotting. Lanes 1-4: HL-60 cells were treated with 0, 1.25, 2.5, and 5.0 μM lycorine, respectively. (D) Subcellular distribution of NF-κB detected by immunofluorescence staining. HL-60 cells were treated with 0 μM (D1, D2, D3) and 5.0 μM lycorine (D4, D5, D6), respectively. Cells were labeled for NF-κB (red) and DNA (blue). Cell nuclei were stained by DAPI (D2, D5). NF-κB was mainly distributed in the cytoplasm (D4, D6) in the HL-60 cells treated with lycorine, whereas NF-κB was mainly distributed in the nucleus in untreated cells (D1, D3). Results were presented as mean ± S.D. (n = 3, three independent experiments). Asterisk symbol (*) indicates significant difference (p < 0.05) compared with the control group.