Fig. 5: DHHC3 palmitoylates TLR9 in the Golgi.
From: Cyclical palmitoylation regulates TLR9 signalling and systemic autoimmunity in mice
A ABE assays were performed in 293 T cells transfected with indicated murine DHHCs and mTLR9-FLAG. A representative blot (left) and mTLR9 relative palmitoylation levels (right, quantified as a ratio of mTLR9-FLAG signal compared to empty vector) are shown (n = 3 replicates). B ABE assays were performed in 293 T cells transfected with murine DHHC3 wild-type (WT) or C157S mutant, DHHC17 WT or C467S mutant, DHHC18 WT or C214S mutant, DHHC20 WT or C214S mutant, along with mTLR9-FLAG. A representative blot (left) and mTLR9 relative palmitoylation levels (right, quantified as a ratio of WT DHHC signal compared to respective mutants) are shown (n = 3 replicates) C TLR9 palmitoylation in DHHC3-deficient cells. ABE assays were performed on DHHC3-deficient RAW264.7 cells retrovirally transduced with mTLR9-HA. A representative blot (left) and mTLR9 relative palmitoylation levels (right, quantified as a ratio of WT signals compared to KO) are shown (n = 3 replicates). D ABE assays were performed in 293 T cells transfected with murine DHHC3 wild-type (WT) along with mTLR7-FLAG. A representative blot (left) and mTLR7 relative palmitoylation levels (right, quantified as a ratio of mTLR7-FLAG signal compared to empty vector) are shown (n = 3 replicates). E ABE assays were performed in 293 T cells transfected with indicated human DHHCs and hTLR9-FLAG. A representative blot (left) and mTLR9 relative palmitoylation levels (right, quantified as a ratio of hTLR9-FLAG signal compared to empty vector) are shown (n = 3 replicates). All data are pooled from three independent experiments (mean ± SEM, P values were calculated by two-way Student’s t test).
