Figure 5: Blocking Notch signaling resulted in attenuated activation of NF-κB and up-regulation of CYLD. | Scientific Reports

Figure 5: Blocking Notch signaling resulted in attenuated activation of NF-κB and up-regulation of CYLD.

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

Figure 5

(a–c) BMDMs derived from the RBP-J cKO and control mice were stimulated with CM prepared from Hepa1-6 cell culture that had been treated with I/R injury in vitro. Nuclear proteins were extracted and p65 protein level was evaluated by using Western blot with LaminA/C as a reference control, and quantitatively compared between the RBP-J cKO and control group (a). Total RNA was extracted and the mRNA level of CYLD was analyzed by using real-time RT-PCR, with β-actin as a reference control (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 between the RBP-J cKO and control group (c). (d–f) RAW264.7 cells were treated with GSI or DMSO and stimulated simultaneously with CM prepared from Hepa1-6 cells that were subjected to I/R injury in vitro. Nuclear proteins were extracted and p65 protein level was evaluated by using Western blot with LaminA/C as a reference control, and quantitatively compared (d). Total RNA was extracted and the mRNA level of CYLD was analyzed by using real-time RT-PCR, with β-actin as a reference control (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, **P < 0.01.

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