EMBO/soda-nel huggingface dataset:
Calcium flux control by Pacs1-Wdr37 promotes lymphocyte quiescence and lymphoproliferative diseases (2021)
Figure 4

FIGURE 4: Pacs1 deletion induces ER stress, ROS, and heightened sensitivity to oxidative stress.

(A) Immunoblot of ER mass, ER stress, and autophagy markers in Pacs1+/+ and Pacs1-/- splenic B cells that were left unstimulated or stimulated overnight with 5 mcg/ml IgM.

(B) B cells were purified from Pacs1+/+ and Pacs1-/- spleens and OCR was measured using a Seahorse metabolic flux analyzer. OCR after rotenone/Antimycin A treatment was subtracted from baseline OCR to give basal mitochondrial respiratory rate. Data shown are the mean of 7-10 technical replicates from three independent experiments. Error bars indicate mean ± SD. Two-tailed unpaired t test, *P < 0.05.

(C) Determination of cellular ROS in Pacs1+/+ and Pacs1-/- FOB cells with CellRox Green. Quantification shows the mean fluorescence intensity (MFI) from three separate pairs of Pacs1+/+ and Pacs1−/− mice. Two-tailed paired t test, **P < 0.01.

(D) Splenocytes from Pacs1+/+ and Pacs1−/− mice were stained with cell surface antibodies to identify FOB cells and treated with 100 µM H2O2 for 35 minutes. Cells were then labelled with TMRE to monitor MMP by FACS analysis. Low TMRE fluorescence indicated susceptibility to H2O2 treatment. Data are presented as mean ± SD of 3-4 replicates, two-tailed unpaired t test, ***P < 0.001. Results are representative of three independent experiments performed on different Pacs1+/+ and Pacs1−/− pairs.
Calcium flux control by Pacs1-Wdr37 promotes lymphocyte quiescence and lymphoproliferative diseases (2021)
Figure 5

FIGURE 5: Pacs1-/- B cells have reduced IP3R expression

(A) Immunoblot of expression of all three IP3R isoforms and SERCA2 in primary splenic B cells from Pacs1+/+ and Pacs1-/- mice.

(B) Real-time quantitative PCR of IP3R and SERCA2 transcripts in B cells. Data are presented as mean ± SD from three independent Pacs1+/+ and Pacs1-/- pairs of mice.

(C) Pacs1-/- FOB cells were stimulated with 0.625 µM thapsigargin (black arrowhead) under Ca2+-free conditions to measure intracellular Ca2+ stores. Kinetic traces of four independent experiments are shown (Pacs1+/+ gray, Pacs1-/- pink) with the mean overlaid in bold (Pacs1+/+ black, Pacs1-/- red).

(D) Plateau of cytosolic Ca2+ flux from intracellular Ca2+ stores in (A) calculated by the mean value over the last 30 seconds of analysis with baseline subtracted. Two-tailed paired t test, *P < 0.05.

(E) AUC of cytosolic Ca2+ flux from intracellular Ca2+ stores in (A). Two-tailed paired t test, *P < 0.05.
Calcium flux control by Pacs1-Wdr37 promotes lymphocyte quiescence and lymphoproliferative diseases (2021)
Figure 6

Figure 6: Pacs1 deletion warps ER Ca2+ handling.

(A) Immunoblot of Pacs1, Wdr37, IP3Rs, and ER stress markers in the parental NIH-3T3 cell line and three separate Pacs1-/- clones.

(B) Real-time quantitative PCR of IP3R isoform expression WT and Pacs1-/- 3T3 cells. Expression in the Pacs1-/- cells was measured in three independent clones. Data are presented as mean ± SD.

(C) Pacs1+/+ and Pacs1-/- NIH-3T3 cells were transfected with cytosolic aequorin and Ca2+ flux was measured after treatment with 1 µM bradykinin. Representative kinetic tracing is shown.

(D) Peak cytosolic Ca2+ concentration based on aequorin measurements as performed in (C). Symbols represent individual wells containing ~5 x 105 transfected 3T3 or Pacs1-/- cells from one experiment. Horizontal bars indicate mean. Two-tailed unpaired t test, **P < 0.01.

(E) Pacs1+/+ and Pacs1-/- NIH-3T3 cells (C1 and C2 from (A)) were transfected with ER-GCamP6-210. ER Ca2+ was measured before and after treatment with 10 µM ATP. Kinetic traces show the mean 488/405 nm excitation ratio of each cell line with error bars indicating SEM. N=30 cells (3T3), 16 cells (C1), 25 cells (C2). Data are representative of 2 independent experiments.

(F) ER Ca2+ release from the NIH-3T3 cell lines imaged in (E). Symbols represent individual cells. Horizontal bars indicate mean value. One-way ANOVA with Tukey's test for multiple comparisons, **P < 0.01, ***P < 0.001.
Calcium flux control by Pacs1-Wdr37 promotes lymphocyte quiescence and lymphoproliferative diseases (2021)
Figure 7

FIGURE 7: Spontaneous proliferation and increased cell death of Pacs1−/− B cells in vivo under lymphocyte replete conditions

(A) Pacs1+/+ and Pacs1−/− B cells were purified, labeled with CTV dye, and stimulated with the indicated mitogens. Cell proliferation was assessed after 72 h with FACS analysis based on CTV dilution.

(B, C) Pacs1+/+ and Pacs1−/− mice were immunized with alum-ova and one week later with NP-Ficoll. Anti-ova IgG and anti-NP IgM titers were measured at 14 days and 7 days after immunization, respectively. Each symbol represents an individual mouse.

(D) Pacs1+/+ and Pacs1ccy/ccy mice were immunized with NP-KLH. Low affinity (anti-NP30) and high affinity (anti-NP2) antibodies were measured 14 days after immunization.

(E) B cells were purified from Pacs1+/+ and Pacs1−/− mice and labeled with CTFR and CTV dyes, respectively. Labeled B cells were injected into unirradiated CD45.1 recipients at ~1:1 ratio. Proliferation and survival of adoptively transferred B cells were measured 8 days post-transplant.

(F) Fraction of donor B cells that proliferated after adoptive transfer from independent experiments using three different Pacs1+/+ and Pacs1−/− donor pairs. Each symbol represents an individual recipient. Horizontal bars indicate median value. Two-tailed unpaired t test, **P < 0.01, ***P < 0.001.

(G) Fraction of donor B cells that were Annexin V positive after adoptive transfer from two independent experiments using two different Pacs1+/+ and Pacs1−/− donor pairs. Each symbol represents an individual recipient. Horizontal bars indicate median value. Two-tailed unpaired t test, **P < 0.01, ***P < 0.001.

(H, I) Pacs1+/+ and Pacs1-/- mice were injected with EdU and the fraction of EdU+ FOB and MZB cells were measured in the spleen at 1, 4, and 7 days post-injection. Data are from one experiment.
Calcium flux control by Pacs1-Wdr37 promotes lymphocyte quiescence and lymphoproliferative diseases (2021)
Figure 8

FIGURE 8: Pacs1 deletion suppresses abnormal lymphocyte accumulation in models of lymphoproliferation

(A) Spleen size and FACS analysis of abnormally expanded B220+CD23+CD21+/low FOB cells in Pacs1+/−;Bcl2TG and Pacs1−/−;Bcl2TG mice.

(B) Number of circulating B cells in the blood and FOB cells in the spleen of Pacs1+/−;Bcl2TG and Pacs1−/−;Bcl2TG mice. Horizontal bars indicate mean value. Two-tailed Mann-Whitney test, *P < 0.05, **P < 0.01.

(C) B cells were purified from the spleens of Pacs1+/−;Bcl2TG and Pacs1−/−;Bcl2TG mice (CD45.2), labelled with CTFR and CTV proliferation dyes, respectively, and transplanted into unirradiated CD45.1 recipients. Donor B cells were measured in the spleen of recipient mice 7 days after B cell transfer based on CD45.2 expression and proliferation dye fluorescence.

(D, E) The fraction of proliferating (D) and recovered (E) donor cells from the experiment in (C). Symbols represent individual recipient mice and data are from two independent adoptive transfer experiments. Two-tailed unpaired t test, **P < 0.01, ***P < 0.001.

(F) Fraction of apoptotic B cells in the adoptively transferred B cell populations in the experiment in (C). Symbols represent individual recipient mice and data are from one adoptive transfer experiment. Two-tailed unpaired t test, ***P < 0.001.

(G) Splenocytes from Pacs1+/−;Bcl2TG and Pacs1−/−;Bcl2TG mice were stained with cell surface antibodies to identify FOB cells and treated with 100 µM H2O2 for 35 minutes. Cells were then labelled with TMRE to monitor MMP. TMRE fluorescence was measured by FACS analysis. Data are presented as mean ± SD of 3-4 replicates, unpaired t test, ns = not significant, ***P < 0.001. Results are from one experiment.

(H) Lymph node size and flow cytometry of lymphoproliferative CD3+B220+ cells in Pacs1+/+;Faslpr/lpr­ and Pacs1−/−;Faslpr/lpr mice.

(I) Enumeration of CD3+B220+ cells in the peripheral blood and lymph nodes of Faslpr/lpr dependent on Pacs1 expression. Horizontal bars indicate median value. Two-tailed Mann-Whitney test, **P < 0.01.

EXPANDED VIEW FIGURE LEGENDS
Structural basis for feedforward control in the PINK1/parkin pathway (2022)
Figure 2

Figure 2. Activation of parkin by pUbl or pUb binding.

A. Ubiquitin-vinyl sulfone (UbVS) assays of RING2 release by addition of pUbl and pUb in trans to R0RBR parkin. Assays were performed with incubation of 2 μM wild-type or W403A R0RBR purified in a 1:1 complex with pUb. Crosslinking was initiated by addition of 10 μM UbVS in the presence of pUbl or pUb∆G76.

B. Inhibition of R0RBR activation by the K211N mutation.

C. UbVS assays of RING2 release by various type of pUb.

D. Autoubiquitination assays of parkin ligase activity induced by pUbl or pUb∆G76 in trans. 2 µM GST-R0RBR in complex with pUb was incubated with 50 nM E1, 2 µM UbcH7, 100 µM ubiquitin and 4 mM ATP.

E. UbVS assays of RING2 release with R0RBR purified without pUb bound.

F. Autoubiquitination assays of parkin ligase activity with GST-R0RBR without pUb bound. The percentage of unmodified parkin band in each lane is indicated under the gel.
Structural basis for feedforward control in the PINK1/parkin pathway (2022)
Figure 3

Figure 3. Ubiquitination activity of Ub-R0RBR chimera.

A. Schematic representations of phosphorylated parkin (pParkin) and the phosphorylated chimera (pUb-R0RBR) with the A320R mutation.

B. Phosphorylation of wild-type (WT) parkin and chimeric Ub-R0RBR by PINK1. Reactions were analyzed on a Phos-tag SDS-PAGE gel to assess the level of phosphorylation.

C. Release of RING2 by Ub-R0RBR chimera. Ubiquitin-vinyl sulfone assays (10 μM UbVS) were performed with 3 μM phosphorylated and non-phosphorylated full-length parkin and Ub-R0RBR.

D. Ubiquitination activity of Ub-R0RBR chimera. Autoubiquitination assays of phosphorylated full-length parkin and Ub-R0RBR (3.3 µM with 3 µm UbcH7 and 75 µM S65A ubiquitin) were performed to assess the impact of pUbl substitution by pUb on parkin activity.
Structural basis for feedforward control in the PINK1/parkin pathway (2022)
Figure 4

Figure 4. Parkin activation by pUb chains.

A. Model of (pUb)4 chain binding to R0RBR.

B. UbVS assays of release of RING2 by K6- or K48-linked (pUb)4 chains. 10 μM UbVS was added to 2 μM R0RBR in the presence of increasing amount of K6(pUb)4 or K48(pUb)4 chains. C. Autoubiquitination assays of parkin activation by K6(pUb)4 or K48(pUb)4. Assays with 2 µM GST-R0RBR, 50 nM E1, 2 µM UbcH7 and 100 µM ubiquitin were performed in the presence of increasing amount of K6(pUb)4 or K48(pUb)4 chains.

D. Competition between K6(pUb)4 chains and monomeric pUb. UbVs (10 μM) was added to 20 nM of WT and K211N R0RBR in the presence of monomeric pUb and/or K6(pUb)4 chains. Reactions were resolved on SDS-PAGE and visualized with anti-parkin antibody by western blotting. The percentage of unmodified parkin band in each lane is indicated under the gel.
Lysine acetylome profiling uncovers novel histone deacetylase substrate proteins in Arabidopsis (2017)
Figure 4

Figure 4. HDA14 protein localizes to the chloroplasts and mitochondria in Arabidopsis, its activity is dependent on cofactors and can be inhibited by deacetylase inhibitors. (A) GFP localization (green) of the HDA14-GFP fusion construct in Arabidopsis protoplasts (35S:HDA14:GFP) from stable transformants. The mitochondrial marker TMRM is depicted in purple. GFP+TMRM shows the overlay image of 35S:HDA14:GFP and TMRM, AF indicates the chlorophyll autofluorescence and BF the bright-field image of the protoplast. GFP+AF+TMRM represents the overlay image of the three fluorescence channels. Scale bar: 10 µM. (B) Deacetylase activity of the recombinant 6xHis-HDA14 protein using a colorimetric assay. Co2+ and Zn2+ were used as cofactors, apicidin (100 µM) and trichostatin A (5 µM) as deacetylase-inhibitors (n = 5, ±SD).
Lysine acetylome profiling uncovers novel histone deacetylase substrate proteins in Arabidopsis (2017)
Figure 5

Figure 5. Differential lysine acetylation and protein expression in hda14 vs. wild-type leaves under growth light (A, D), in isolated thylakoids (B, E), and under low light conditions (C,F). Volcano plots depict lysine acetylation site ratios (A-C, top row) or protein ratios (D-F, bottom row) for mutant vs. control, with p-values determined using the LIMMA package. Orange, protein with nuclear localization; Green, protein with plastidial localization; purple triangles, proteins of the Calvin-Benson (CB) cycle; blue diamonds (top row), proteins of the light reaction; localization information according to SUBA4 database. Blue circles (bottom row), proteins with lysine acetylation sites identified. Dashed lines indicate significance thresholds of either uncorrected p-values < 5% or Benjamini-Hochberg corrected FDR < 5%. A missing FDR line indicates that the 5% threshold was not reached by any of the data points.
Lysine acetylome profiling uncovers novel histone deacetylase substrate proteins in Arabidopsis (2017)
Figure 6

Figure 6. RuBisCO activity and RuBisCO activation state is increased in the hda14 mutant under low light conditions. (A) RuBisCO initial and total activity in WT and hda14 in low light treated plants. Initial activity was measured directly upon extraction. For the total activity samples were incubated with H2CO3 for 3 minutes to fully carbamylate the active site of RuBisCO (n = 10, *t-test p-value < 0.05). (B) RuBisCO activation state. (C) ATPase activity of recombinant 6x-HisRCAß1 WT, K438Q and K438R with ATP and ADP/ATP= 0.11, respectively (n = 3, *t-test p-value < 0.05, +t-test p-value< 0.1).
Cellular stress promotes NOD1/2-dependent inflammation via the endogenous metabolite sphingosine-1-phosphate (2021)
Figure 1

Fig.1 NOD1/2 sense perturbation of cellular homeostasis

(A, B) qRT-PCR analysis of IL6 expression upon indicated stimulations in presence or absence of NOD1 (A) or NOD2 (B). HeLa inducible NOD1 cells (A) or NOD2 cells (B) were induced in absence or presence of doxycycline overnight and afterwards stimulated with various stimuli for 4 h.

(C, D) ELISA analysis of IL6 production in supernatants of HeLa NOD1 cells (C) or NOD2 cells (D) upon different stimulations. HeLa NOD1 cells (C) or NOD2 cells (D) were induced in absence or presence of doxycycline overnight, stimulated with various stimuli for 20 h and afterwards supernatants were collected for ELISA.

(E, F) Samples mean value of Eigengene analysis of TNFα signaling via NF-κB pathway (E) and inflammatory response pathway (F) of HeLa NOD1 or NOD2 cells upon indicated stimulations.

Data information: (A, B) Means±SD of 3 technical replicates from one representative experiment out of 4 independent experiments. P values were calculated with ANOVA test of linear mixed model for fix effect of main factors. (C, D) Means±SEM of 3 independent experiments. Each dot represents one independent experiment. P values were calculated using two-way ANOVA. (*) p ≤0.05, (**) p ≤ 0.01, (***) p ≤ 0.001, (****) p ≤ 0.0001.
Cellular stress promotes NOD1/2-dependent inflammation via the endogenous metabolite sphingosine-1-phosphate (2021)
Figure 2

Fig. 2 NOD1/2 signaling is required for NF-κB activation and inflammatory response upon perturbation of cellular homeostasis.

(A) NF-κB luciferase assay in HEK293T wildtype (WT) and NOD1/2 knockout (KO) NF-κB reporter cells upon various stimulations.

(B) qRT-PCR analysis of IL8 expression upon indicated stimulations in HEK293T WT and NOD1/2 KO cells.

(C) qRT-PCR analysis of Cxcl2 expression upon indicated stimulations in WT, Nod1/2 KO and Rip2 KO bone marrow derived macrophages (BMDMs).

(D) Multiplex analysis of CXCL2 production in supernatants of WT, Nod1/2 and Rip2 KO BMDMs upon indicated stimuli for 20 h.

Data information: (A-C) Means±SD of 3 technical replicates from one representative experiment out of 3 independent experiments. (D) Means±SEM of 3 independent experiments. Each dot represents one independent experiment. P values were calculated using two-way ANOVA. (*) p ≤0.05, (**) p ≤ 0.01, (***) p ≤ 0.001, (****) p ≤ 0.0001.
Cellular stress promotes NOD1/2-dependent inflammation via the endogenous metabolite sphingosine-1-phosphate (2021)
Figure 3

Fig. 3 S1P generation is essential for pro-inflammatory responses induced by stressors and by Shigella infection

(A) Fluidigm analysis of expression of genes involved in sphingolipid metabolism in human dermal fibroblasts (HDFs) upon indicated stimulations for 4 h. Heatmap shows means of 4 independent experiments. The color scale represents the relative gene expression compared to control from lower (blue) to higher levels (red).

(B) Lipidomic profiling of sphingolipid metabolites in HDFs upon indicated stimulations for 2 h. Fold changes of each metabolite were calculated against corresponding controls. Heatmap shows means of 3 independent experiments. The color scale represents the relative levels of various lipids from lower (blue) to higher abundance (red).

(C) ELISA analysis of cytosolic S1P production in HDFs upon indicated stimulation for 4 h.

(D) qRT-PCR analysis of IL6 expression in HDFs upon indicated stimulations for 4 h in absence or presence of the inhibitor of sphingosine kinases-SKI II (SPHKi, 50 µM).

(E-G) ELISA analysis of IL6 in supernatants of HDFs (E), HeLa NOD1 cells (F) or HeLa NOD2 cells (G) upon indicated stimulations for 20 h in absence or presence of the inhibitor of sphingosine kinases.

(H) qRT-PCR analysis of Cxcl2 expression in WT, Sphk1 KO or Sphk2 KO BMDMs upon indicated stimulations for 4 h.

(I) Multiplex analysis of CXCL2 production in supernatants of WT, Sphk1 and Sphk2 KO BMDMs upon indicated stimulations for 20 h.

(J) ELISA analysis of IL8 production upon Shigella infection. HeLa scrambled control cells and SPHK1/2 double KD cells were infected with Shigella flexneri M90T for 3 h and 6 h, or stimulated with TNFα for 6 h.

Data information: (C, E-G, I, J) Means±SEM of 3 independent experiments. Each dot represents one independent experiment. (D, H) Means±SD of 3 technical replicates from one representative experiment out of 3 independent experiments. P values were calculated using one-way or two-way ANOVA. (*) p ≤0.05, (**) p ≤ 0.01, (***) p ≤ 0.001, (****) p ≤ 0.0001.
Cellular stress promotes NOD1/2-dependent inflammation via the endogenous metabolite sphingosine-1-phosphate (2021)
Figure 4

Fig. 4 Cytosolic S1P induced NF-κB activation and inflammation is mediated by NOD1/2

(A, B) ELISA analysis of IL6 in HeLa NOD1 cells (A) or HeLa NOD2 cells (B) upon indicated stimulations for 20 h. For cytosolic delivery of S1P, iE-DAP or MDP, cells were treated together with digitonin (5 µg/ml).

(C) GFP immunoprecipitation (IP) of NOD1/2 upon S1P, iE-DAP or MDP stimulation. S1P (20 µM), iE-DAP (20 µM) or MDP (20 µM) were delivered into the cytosol of HeLa NOD1 or HeLa NOD2 cells with digitonin (5 µg/ml) for 1 h. Afterwards cell lysates were collected for IP.

(D) Western blot analysis of MAPKs and NF-κB activation upon S1P stimulation. HeLa NOD1 or NOD2 cells were treated with cytosolic S1P (20 µM), iE-DAP (20 µM) or MDP (20 µM) for indicated times.

(E) NF-κB luciferase assay in HEK293T WT and NOD1/2 KO NF-κB reporter cells upon cytosolic S1P stimulation.

(F) qRT-PCR analysis of IL8 expression upon cytosolic S1P stimulation in HEK293T WT and NOD1/2 KO cells.

(G) qRT-PCR analysis of Il6 expression in WT, Nod1/2 and Rip2 KO BMDMs upon S1P (10 µM) or MDP (10 µM) stimulations together with digitonin (2.5 µg/ml) for 4 h.

(H-K) Multiplex analysis of IL6 (H), CXCL2 (I), CCL5 (J) and IL16 (K) production in supernatants of WT, Nod1/2 and Rip2 KO BMDMs upon S1P (10 µM) or MDP (10 µM) stimulations together with digitonin (2.5 µg/ml) for 20 h.

Data information: (A, B, E-G) Means±SD of 3 independent experiments. (H-K) Means±SEM of 3 independent experiments. Each dot represents one independent experiment. P values were calculated using two-way ANOVA (A, B, E, G-K) or Student's t test (F). (*) p ≤0.05, (**) p ≤ 0.01, (***) p ≤ 0.001, (****) p ≤ 0.0001.
Cellular stress promotes NOD1/2-dependent inflammation via the endogenous metabolite sphingosine-1-phosphate (2021)
Figure 5

Fig. 5 S1P interacts with NOD1/2

(A, B) Immunoprecipitation with various lipid-coated beads using HeLa NOD1-GFP (A) and NOD2-GFP cells (B). Cell lysates of HeLa NOD1 or HeLa NOD2 cells were incubated with lipid-coated beads at room temperature for 2 h.

(C) Immunoprecipitation with S1P-coated beads using THP-1 lysates.

(D) Dual-color dSTORM imaging of HeLa NOD1 or NOD2 upon S1P stimulation. After stimulation with S1P-TAMRA (20 µM) for 2 h, HeLa NOD1 or NOD2 cells were fixed, permeabilized and stained with anti-GFP antibody and Alex Fluor 647-conjugated secondary antibody. Images were reconstructed from 10,000 raw frames. Scale bar: 200 nm.

(E, F) Single particle tracking of NOD1-GFP (E) or NOD2-GFP (F) with S1P-TAMRA. After doxycycline induction overnight, HeLa NOD1 or NOD2 cells were stimulated with S1P-TAMRA (20 µM) for 1 h together with digitonin. Scale bar: 5 µm.
Cellular stress promotes NOD1/2-dependent inflammation via the endogenous metabolite sphingosine-1-phosphate (2021)
Figure 6

Fig. 6 S1P directly binds to NOD1/2 via the NBD domain and triggers their activation

(A) Molecular docking of S1P with rabbit NOD2.

(B) SDS-PAGE analysis of purified NOD1-GFP and NOD2-GFP.

(C) Microscale thermophoresis (MST) analysis of direct binding of S1P to NOD1, NOD2, NLRP3 or GFP.

(D) MST analysis of direct binding of NOD1 to S1P and iE-DAP.

(E) MST analysis of direct binding of NOD2 with S1P and MDP.

(F) ELISA analysis of IL8 in supernatants of HEK293T NOD1/2 KO cells transfected with indicated NOD1 or NOD2 mutants. After 24 h transfection, cells were stimulated with S1P (20 µM), iE-DAP (20 µM) or MDP (20 µM) together with digitonin for 20 h.

(G, H) MST analysis of direct binding of NOD1 (G) or NOD2 (H) with iE-DAP or MDP in the presence of S1P (2 µM).

Data information: (C) Means±SD from 3 independent experiment. (D, E, G, H) One representative experiment out of 3 independent experiments. (F) Means±SD of 3 technical replicates from one representative experiment out of 3 independent experiments. P values were calculated using two-way ANOVA. (*) p ≤0.05, (****) p ≤ 0.0001.
Cellular stress promotes NOD1/2-dependent inflammation via the endogenous metabolite sphingosine-1-phosphate (2021)
Figure 7

Fig. 7. S1P binds to NOD1 via H517 and impairs ADP binding.

(A, B) MST analysis of direct binding of NOD1 (A) or NOD2 (B) with ADP in the presence of S1P (2 µM).

(C, D) MST analysis of direct binding of NOD1 (C) or NOD2 (D) with S1P in the presence of ADP (2 µM).

(E, F) MST analysis of direct binding of S1P (E) or ADP (F) with NOD1 WT or NOD1 H517A.

Data information: (A-F) One representative experiment out of 3 independent experiments.

Figure legends