Fig. 6: Degradation of DUX4 during HSV-1 infection using the mini-TRIMAway assay.
From: Herpesviruses mimic zygotic genome activation to promote viral replication
A Principle of the mini-TRIMAway assay: A DUX4 specific nanobody linked to the TRIM21 ring targets DUX4 for proteosomal degradation. B AlphaFold-Multimer generated prediction of the interaction of the two DUX4 directed nanobody clones Nb2-4 and Nb2-19 to the DUX4 protein. C Western blot analyzing DUX4 levels after simultaneous induction of DUX4 and induction of the nanobody targeting DUX4 for degradation (nb 2–4 or nb 2–19) or control nanobody (nb ctrl). HEK293T were induced with 0,1 µg/ml Doxycycline and analyzed 24 h post induction. HSP90 was used as loading control. D Microscope analysis of the mini-TRIMAway assay for DUX4 degradation after HSV-1 infection at day 1–3 post infection. The corresponding nanobody (nb 2-4, nb 2-19 or nb ctrl) was induced with 2 µg/ml doxycycline and infected with HSV-1 GFP (MOI 0.05). Representative experiment out of n = 3. E Measurement of GFP-expression by flow cytometry of the mini-TRIMAway assay for DUX4 degradation after HSV-1 infection at day 1-3 post infection. The corresponding nanobody (nb 2–4, nb 2–19 or nb ctrl) was induced with 2 µg/ml doxycycline and infected with HSV-1 GFP (MOI 0.05). Representative experiment out of n = 3. Values shown are technical replicates. Source Data are provided as a Source Data file.
