c-Myb Dominates TBK1-Mediated Endotoxin Tolerance in Kupffer Cells by Negatively Regulating DTX4

Figure 4

The transcription factor c-Myb can negatively regulate DTX4 to induce the K48-linked polyubiquitination profile of TBK1: (a) the interaction between TBK1 and DTX4 in KCs was verified by immunoprecipitation (IP); (b) the protein expression of NLRP4, c-Myb, and DTX4 in KCs was detected by Western blotting. Densitometric analysis of the Western blotting data; (c) position weight matrix of transcription factor c-Myb binding site (right panel). The c-Myb binding sites and the first three binding sequences of the DTX4 promoter regions were obtained from the JASPAR website (left panel); (d) the ChIP assay was performed in HEK293T cells to verify the three binding sequences of c-Myb in the DTX4 promoter region; (e) luciferase reporter assay was performed in HEK293T cells to verify that transcription factor c-Myb could negatively regulate DTX4; (f) the regulation of c-Myb on DTX4 in RAW264.7 cells was detected by Western blotting; (g) K48-linked polyubiquitination of TBK1 in RAW264.7 cells was detected. Data are represented as the mean ± SD of at least three independent experiments; one-way ANOVA with Dunnett’s multiple comparisons was used. , .

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