Fig. 5

PKC-412 activates the NF-kappa B signaling pathway. The 293 T cells (a), A3.01 cells (b), and resting PBMCs (d) were transduced by Cignal Lenti particles encoding a luciferase (luc) gene under the control of a minimal (m)CMV promoter (Promo) and tandem repeats of the NF-κB transcriptional response element (TRE). After being transduced, cells were treated with different concentrations of PKC412 and TNF-α for 12 h and activation of NF-κB signaling was detected by measuring the luc activity. d ACH2 cells were treated with various concentrations of PKC412 and TNF-α for 12 h, and then cells were fractionated into nuclear and cytoplasmic fractions. The nucleus- and cytoplasm-associated p65, histone-4 and β-tubulin were detected by WB with corresponding antibodies. e ACH2 cells were treated with various concentrations of PKC412 and TNF-α for 12 h. Then the cells were lysed, and Ser³¹¹phosphorylated p65 was detected by WB with corresponding antibody. Error bars represent variations between triplicate samples and the data are representative of results obtained in three independent experiments. The degree of significance for the PKC412 or TNF-α treatment was reported relative to the mock treatment. ** p < 0.01, *** p < 0.001, and **** p < 0.0001