Fig. 3: The effect of USP29 on TAK1 ubiquitination and phosphorylation. | Cell Death Discovery

Fig. 3: The effect of USP29 on TAK1 ubiquitination and phosphorylation.

From: ELAVL1-mediated USP29 mRNA degradation activates TAK1 driving M1 microglial polarization and neural stem cell differentiation dysregulation in spinal cord injury

Fig. 3

A Network diagram of USP29 and known deubiquitination substrates from the UbiBrowser 2.0 database. B Venn diagram of USP29 substrates and genes related to SCI and microglial polarization. C Western blot analysis of p-TAK1 and TAK1 protein expression in spinal cord tissues of rats on day 7 after SCI (N = 6). D Western blot analysis of p-TAK1 and TAK1 protein expression in HMC3 cells of each group. E RT-qPCR analysis of USP29 expression in HMC3 cells. F Western blot analysis of p-TAK1 and TAK1 protein expression in HMC3 cells of each group. G Western blot analysis of USP29, p-TAK1, and TAK1 protein expression in HEK293T cells transfected with different doses of oe-USP29 plasmids (0, 0.5, 1, or 2 μg) for 48 h. H CO-IP analysis of USP29 interaction with TAK1 in HEK293T cells co-transfected with Flag-USP29 and Myc-TAK1 plasmids. I GST pull-down experiment assessing the direct interaction between USP29 and TAK1 in HEK293T cells, with GST-Myc plasmid as a control. J After transfecting HEK293T cells with Myc-TAK1, Flag-USP29, and HA-Ub plasmids for 48 h, immunoprecipitation was performed using anti-Myc agarose beads, followed by Western blotting with anti-HA and anti-Myc antibodies. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. All cell experiments were repeated three times.

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