Fig. 4

UBASH3A inhibits phosphorylation of NF-κB dependent on TCR signaling. A, B CD4⁺ T cells were transfected with pcDNA3.1-UBASH3A and empty vector. A At 24 h after transfection, UBASH3A mRNA levels were quantified by RT-qPCR. The amount of UBASH3A transcript was expressed relative to that of the GAPDH transcript. B At 30 min after TCR stimulation, Western blotting was performed with a series of antibodies. Data are representative of three independent Western blotting experiments. C The amount of total (t)-NF-κB was normalized to that of β-actin and that of phosphorylated (p)-NF-κB was then normalized to that of normalized t-NF-κB. Results were plotted on a histogram. Values are expressed as mean ± standard deviation. *P<0.05, vs lane 1 or 4, by Dunnett’s multiple comparison test. RQ, relative quantification