Figure 4: Myeloid-specific RBP-J deletion reduced the production of TNF-α and IL-1β by macrophages treated with conditional medium derived from I/R-injured hepatocytes in vitro. | Scientific Reports

Figure 4: Myeloid-specific RBP-J deletion reduced the production of TNF-α and IL-1β by macrophages treated with conditional medium derived from I/R-injured hepatocytes in vitro.

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

Figure 4

(a–c) BMDMs were generated from RBP-J cKO and control BM monocytes, and stimulated 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). Total RNA was extracted from the RBP-J cKO and control BMDMs stimulated with CM, and the mRNA level of TNF-α and IL-1β was examined by using real-time RT-PCR, with β-actin as a reference control (b). The level of ROS in the RBP-J cKO and control BMDMs was examined by using FACS, and was quantified with MFI (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. The protein level of TNF-α and IL-1β in the supernatant was determined by using ELISA (d). Total RNA was extracted from the GSI- or DMSO-treated BMDMs stimulated with CM, and the mRNA level of TNF-α and IL-1β was examined by using real-time RT-PCR, with β-actin as a reference control (e). The level of ROS in the GSI- or DMSO-treated BMDMs was examined by using FACS, and was quantified with MFI (f). Bars = mean ± SD (n = 5). *P < 0.05, **P < 0.01, ns, not significant.

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