Figure 7: Forced Notch activation led to increased production of TNF-α and IL-1β and increased activation of NF-κB in macrophages suffering from I/R injury in vitro. | Scientific Reports

Figure 7: Forced Notch activation led to increased production of TNF-α and IL-1β and increased activation of NF-κB in macrophages suffering from I/R injury in vitro.

From: Blocking Notch signal in myeloid cells alleviates hepatic ischemia reperfusion injury by repressing the activation of NF-κB through CYLD

Figure 7

(a–c) BMDMs were co-cultured with OP9-Dll1 (Dll1) or OP9-GFP (Ctrl) cells, and treated with CM prepared from Hepa1-6 cells that were subjected to I/R injury in vitro. The protein level of TNF-α and IL-1β in the supernatant was determined by using ELISA (a). Nuclear proteins were extracted and p65 protein level was evaluated by using Western blot with LaminA/C as a reference control, and quantitatively compared (b). Total proteins were extracted and CYLD protein level was evaluated by using Western blot with β-actin as a reference control, and quantitatively compared (c). (d,e) RAW264.7 cells were co-cultured with OP9-Dll1 or OP9-GFP cells, and were treated with CM prepared from Hepa1-6 cells that were subjected to I/R injury in vitro. The protein level of TNF-α and IL-1β in the supernatant was determined by using ELISA (d). Nuclear proteins were extracted and p65 protein level was evaluated by using Western blot with LaminA/C as a reference control, and quantitatively compared (e). Total proteins were extracted and CYLD protein level was evaluated by using Western blot with β-actin as a reference control, and quantitatively compared (f). Bars = mean ± SD (n = 5), *P < 0.05.

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