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. 2022 Jul 21;36:44–51. doi: 10.1016/j.jot.2022.07.001

Figure 5.

Figure 5

AptTNF-α regulated TNF-α-mediated necroptosis via RIP1/RIP3/MLKL signalling pathway. A,B,C, the mRNA levels of RIPK1, RIPK3 and MLKL gene were measured by RT-PCR after incubation with TNF-α and AptTNF-α for 24 ​h (n ​= ​3, each group was repeated from three batches of samples for analysis). ∗p ​< ​0.05. D, Expression level of RIPK1, RIPK3 and MLKL, along with their phosphorylated proteins in the BMECs examined by the Western blotting. E,F,G,H, the relative protein level of GAPDH, p-RIP1/RIP1, p-RIP3/RIP3 and p-MLKL/MLKL from the Western blotting analysis, respectively. The ratio of p-RIP1/RIP1, p-RIP3/RIP3 and p-MLKL/MLKL was normalized to GAPDH (n ​= ​3, each group was repeated from three batches of samples for analysis). ∗p ​< ​0.05. BMEC, bone microvascular endothelial cell. AptTNF-α, TNF-α aptamer. Apt 100, 100 ​nM TNF-α aptamer. Apt 500, 500 ​nM TNF-α aptamer.