Extended Data Fig. 8: AMFR mediates K27-linked polyubiquitination of TAB3 on Lys649, but not regulates the degradation of TAB3. | Nature Microbiology

Extended Data Fig. 8: AMFR mediates K27-linked polyubiquitination of TAB3 on Lys649, but not regulates the degradation of TAB3.

From: Staphylococcal virulence factor HlgB targets the endoplasmic-reticulum-resident E3 ubiquitin ligase AMFR to promote pneumonia

Extended Data Fig. 8

(a) Immunoblot analysis of ubiquitination of TAB3 in HEK293T cells co-transfected with MYC-TAB3 mutant (K649R) and FLAG-AMFR, along with the mutant ubiquitin HA-Ub (K63). (b) Luciferase assay of NF-κB activation in HEK293T cells transfected with indicated plasmids. (c) Immunoblot analysis of the expression of TAB3 in cell lysates of HEK293T cells co-transfected with MYC-TAB3 along with increasing amount of FLAG-AMFR for 48 h. Before harvest, cells were treated with or without proteasome inhibitor MG-132 (25 mM) or lysosomal inhibitor (NH4CL 10 mM or leupeptin 20 mM) for 6 h. Densitometric analysis of immunoblot analysis was shown below. (d) Coimmunoprecipitation and immunoblot analysis of HEK293T cells transfected with indicated plasmids. Data shown are representative of three independent experiments, or presented as means ± SD, n = 3 biological replicates, two-way ANOVA (Sidak’s test).

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