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. 2014 May 14;111(21):7683–7688. doi: 10.1073/pnas.1310972111

Fig. 3.

Fig. 3.

PKD1 regulates IL-6/IL-8 at mRNA level and depends on NF-κB activity in Ras OIS. (A) PKD1 regulates IL-6 at mRNA level. Total mRNA were extracted from ER:Ras IMR90 cells expressing indicated genes and shRNAs after 6 d Ras induction. Relative mRNA level of IL-6 was determined by quantitative PCR. mRNA levels in young IMR90 cells are considered as control. (B) PKD1 affects NF-κB binding activity to the promoter of IL-6 gene. ER:Ras IMR90 cells expressing indicated genes and shRNAs were given 4-OHT for 6 d, and then the lysates were analyzed by ChIP assay using an antibody against NF-κB. NF-κB binding to the IL-6 promoters in the indicated stable transfectants is represented relative to mouse IgG binding and is normalized to young IMR90 cells binding for each other. Error bars represent means + SD (n = 3) *P < 0.05, **P < 0.01 in A and B. (C) PKD1 affects IκBα degradation. ER:Ras IMR90 cells expressing WZL-PKD1, WZL-PKD1-DN, and miR30-PKD1, respectively, were given 4-OHT for the indicated times, and then subjected to Western blot to detect IκBα protein levels. (D) NF-κB inhibition abolishes PKD1-mediated IL-6/IL-8 expression. ER:Ras IMR90 cells expressing WZL-PKD1 were treated with solvent DMSO or IκB kinases inhibitor BAY 11-7082 (5 μM) in the presence of 4-OHT for the indicated days, then analyzed for expression of indicated proteins.