The effect of TNF-α on AQP3 protein and plasma membrane water permeability in DJM-1 cells. (A) DJM-1
amyloid A protein fragment were treated with 20 ng/ml TNF-α for 0, 2, 6, 12 or 24 h. Twenty micrograms of cell lysates were collected for Western blotting, and AQP3 was detected with anti-AQP3 antibody. β-Actin was detected with anti-β-actin antibody as an internal control. (B) Cells were treated with 0.02, 0.2, 2 or 20 ng/ml TNF-α for 24 h. AQP3 protein expression was detected by Western blotting. (C) DJM-1 cells were treated with 20 ng/ml TNF-α for 24 h. Plasma membrane water permeability was measured using the stopped-flow method. Relative water permeability was calculated from four different experiments. Each data point represents the mean ± SE. ?p
cells were transfected with 100 nM small interfering RNAs for TNFR1 and incubated for 24 h. Subsequently, cells were treated with 20 ng/ml TNF-α for 24 h. AQP3 protein expression was detected by Western blotting.
