Fig. 3.

UFMylation regulates responses to IFN-γ and LPS. (A) The Ufm1 conjugation cascade. (B) Western immunoblot for free and conjugated Ufm1 in WT and ΔUfm1 (separate run) and ΔUfsp2/ΔUba5/ΔUfc1 cells and cells reconstituted with WT protein or functionally inactive mutant protein or the UFIM mutant for Uba5. Total protein per lane is shown. (C–E) Average IFN-γ–dependent NO production in knockout and complemented lines. (F–H) Average LPS-dependent NO production in knockout and complemented lines. Average data pooled from four to five independent experiments are shown as mean ± SEM relative to knockout cells expressing WT protein stimulated with 10 U IFNγ/mL or 50 ng LPS/mL. *P < 0.05 determined by ANOVA with Tukey’s multiple comparison test; ns, not statistically significant.