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Figure 4 | BMC Cancer

Figure 4

From: Regulation of HSP27 on NF-κB pathway activation may be involved in metastatic hepatocellular carcinoma cells apoptosis

Figure 4

HSP27 interacts with key molecular (IKKβ, IKKα, IκBα) of NF-kB pathway in HCC cell lines. The precipitated IKKβ, IKKα, IκBα with anti-HSP27 antibody or normal mouse IgG1 were detected by western blot analysis in Hep3B, MHCC97L and MHCC97H cells, the HSP27 contents in the precipitates were used as equal loading amount control (A). Endogenous IKK activity in Hep3B, MHCC97L and MHCC97H cells (B) and in RNAi, MOCK and MHCC97H cells (D) was detected through IKK assay. The absorbed value of reaction solution from MHCC97H cells or Hep3B cells served as a percent control. The precipitated HSP27 and phosphorylated HSP27 with IKK complex by using anti-IKKα antibody were detected in Hep3B, MHCC97L and MHCC97H cells, the IKKα contents in the precipitates were used as equal loading amount control (C). Moreover, the precipitated IKKβ with anti-IKKα antibody or precipitated IKKα with anti-IKKβ antibody and normal rabbit IgG1 in RNAi, MOCK and MHCC97H cells were demonstrated by western blot analysis and the association between IKKα and IKKβ was reduced. The IKKα or IKKβ contents in the precipitates were used as equal loading amount control (E). In addition, total IκBα (F) and phosphorylated IκBα (G) were also analyzed by western blot analysis or ELISA in RNAi, MOCK and MHCC97H cells. The values on bottom of the bands represent the densitometric estimation of the relative density of the band. Data are representative of 3 separate experiments; * statistical difference from control at P < 0.05.

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