Figures & data
Figure 1. Effects of hyperthermia on TNF-α-induced expression of E-selectin (a) and VCAM-1 (b) in HAEC monolayers. After hyperthermia, HAEC was stimulated with TNF-α at each time point. E-selectin or VCAM-1 expression was measured by ELISA 4 hours or 6 hours after TNF-α stimulation. Data are shown as a mean ± SEM of five samples in a representative experiment. Similar results were obtained in three independent experiments. #p < 0.001 compared with control. *p < 0.001 compared with TNF-α stimulation alone.
Figure 2. The induction of HSP70, HO-1 and IkBα proteins after hyperthermia HAEC was treated with hyperthermia for 60 min. After heat treatment, total cell protein was extracted at the indicated time point. The productions of HSP70, HO-1, and IBα were assessed by Western blotting. Lane 1; control, Lane 2; 3 hours after hyperthermia, Lane 3; 6 hours after hyperthermia, Lane 4; 24 hours after hyperthermia, Lane 5; 48 hours after hyperthermia.
Figure 3. Effects of hyperthermia on TNF-α-induced activation of NF-κB in HAEC. (a) The status of NF-κB activation was evaluated by Western blotting of p65 in nuclear protein. Upper band shows the Western blotting of nuclear p65 and lower band shows the Western blotting of cytoplasmic P65. (b) Quantification of each band was performed by densitometry analysis software (image J), and results were expressed as the ratio (cytoplasm) in arbitrary units. Lane 1; control, Lane 2; hyperthermia alone, Lane 3; TNF-α alone, Lane 4; hyperthermia pretreatment with TNF-α.
Figure 4. Effects of geranylgeranylacetone (GGA) on accumulation of HSP70 after hyperthermia in HAEC. We added GGA (0.01, 0.1, 1.0, 10 μM) 2 hours before hyperthermia, and HSP70 expression was then assessed by Western blotting at 3 h after heat treatment. Lane 1; control, Lane 2; GGA 0.01 μM alone, Lane 3; GGA 0.1 μM alone, Lane 4; GGA 1 μM alone, Lane 5; GGA 1 μM alone, Lane 6; hyperthermia alone, Lane 7; GGA 0.01 μM before hyperthermia, Lane 8; GGA 0.1 μM before hyperthermia, Lane 9; GGA 1 μM before hyperthermia, Lane 10; GGA 1 μM before hyperthermia.
Figure 5. Effects of geranylgeranylacetone (GGA) on inhibition of TNF-α-induced ECAM accumulation by hyperthermia in HAEC. We added GGA (0.01–1.0 μM) just 2 h before hyperthermia. Three hours after heat treatment, we incubated HAEC with recombinant human TNF-α (10 ng/ml). E-selectin (a) or VCAM-1 (b) expression was measured by ELISA 4 hours or 6 hours after TNF-α stimulation. Data are shown as a mean ± SEM of five samples in a representative experiment. Similar results were obtained in three independent experiments. #p < 0.001 compared with control. *p < 0.001 compared with TNF-α combined with hyperthermia.
