Correction to: Scientific Reports 10.1038/s41598-018-31734-6, published online 07 September 2018
This Article contains an error.
As a result of an error during the figure assembly, in Figure 6B, the panels Flag and GAPDH of the “Input” are overlapping with the Flag and GAPDH panels of the “Input” in Figure 6C.
The corrected Figure 6 and its accompanying legend appears below as Figure 1.
Figure 1.
USP4 removes K48-linked polyubiquitination from TRAF6. (A) Western blot analysis of HEK293T cells transfected with Flag-TRAF6, HA-USP4, and HA-Ub for ubiquitination levels; GAPDH was used as a loading control. (B,C) HEK293T cell lysates transfected with Flag-TRAF6, HA-Ub, together with either wild-type HA-USP4 (WT) or mutated HA-USP4 (CA) were collected and immunoprecipitated with anti-Flag agarose beads. The eluted immunocomplexes was then subjected to SDS-PAGE analysis with anti-ubiquitin antibody. (D) Lysates from HEK293T cells transiently expressing Flag-tagged TRAF6 and HA-tagged ubiquitin, together with either HA-tagged USP4 or its truncated constructs (N or C) were immunoprecipitated with anti-Flag agarose beads; the eluted protein complexes was subjected to western blot analysis with anti-ubiquitin antibody. (E) HEK293T cells transfected with Flag-TRAF6, Myc-USP4, together with either WT HA-Ub or its mutants (K48 or K63) were harvested and subjected to immunoprecipitation with anti-Flag agarose beads followed by SDS-PAGE analysis with HA antibody. (F) Lysates from HEK293T cells transfected with HA-USP4 or a control vector followed by treatment with poly (I:C) (HMW; 30 μg/ml) for 8 h were subjected to immunoprecipitation with an anti-TRAF6 antibody. This was followed by western blot analysis of eluted immunocomplexes with K48 linkage-specific ubiquitin antibodies. (G) HEK293T cells transfected with Flag-TRAF6, control siRNA or USP4-specific siRNA, together with either WT HA-Ub or its mutants (K48 or K63) were harvested and subjected to immunoprecipitation with anti-Flag agarose beads followed by SDS-PAGE analysis with anti-ubiquitin antibody. (H) Western blot analysis of RD cells transfected with control plasmid or HA-USP4 plasmid for 48 h, followed by treatment with EV71 (MOI = 0.5) for 12 h. The cell lysates were subjected to immunoprecipitation with anti-TRAF6 antibody, followed by western blot analysis of eluted immunocomplexes with K48 linkage-specific ubiquitin antibodies.