Figure 8.
Paricalcitol does not affect NF-κB activation and its nuclear translocation induced by TNF-α. HKC-8 cells were pretreated with or without 10−7 M paricalcitol for 0.5 h, followed by incubation with 2 ng/ml TNF-α for various periods as indicated. (A) Western blot showed that paricalcitol did not significantly affect p65 NF-κB phosphorylation and activation induced by TNF-α. (B) Paricalcitol also did not significantly alter IκBα phosphorylation and subsequent degradation induced by TNF-α. Cell lysates were immunoblotted with specific antibodies against phospho- and total p65 NF-κB and against phospho- and total IκBα, respectively. The low band in the doublet (B, lane 1) likely represents the unphosphorylated IκBα. (C) Immunofluorescence staining demonstrated that paricalcitol pretreatment did not affect p65 NF-κB to undergo nuclear translocation upon TNF-α stimulation in HKC-8 cells.