Skip to main content
. Author manuscript; available in PMC: 2020 Jun 4.
Published in final edited form as: Structure. 2019 May 2;27(6):1000–1012.e6. doi: 10.1016/j.str.2019.03.022

Figure 4. FAM105A does not counteract LUBAC activity.

Figure 4.

(A) Immunoblot using anti-Met1Ub antibody detected decreased levels of Met1Ub-conjugated proteins in HEK293T cells upon co-expression of FLAG-OTULIN, but not FLAG-OTULINC129S or FAM105A with HAHOIL and myc-HOIP. (B) Immunoprecipitation with anti-HA antibody (IP:anti-HA) of HA-tagged FAM105A (HA-FAM105A) does not recover myc-tagged HOIP (myc-HOIP). In immunoprecipitation with anti-myc antibody (IP: anti-myc), however, myc-HOIP recovered HA-FAM105A. Input amounts and Tubulin levels are shown. (C) Co-expression of HAFAM105A did not affect activity of FLAG-OTULIN in a luciferase reporter assay. FLAG-OTULIN represses myc-HOIP/HA-HOIL dependent activation of an NFκB-dependent luciferase reporter. Co-transfection of HA-FAM105A does not impact myc-HOIP/HA-HOIL dependent activation or FLAG-OTULIN dependent repression of this luciferase reporter. General transcriptional activity of an AP1 luciferase reporter was unaffected. HA-MAPKK1-R4F acts as a positive control for the AP1 luciferase assay.