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d609  (MedChemExpress)


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    MedChemExpress d609
    Increased sEV secretion in cholesterol‐depleted conditions is due to autophagy‐driven signalling. (A) Representative images for CD63 and LC3B expression in HEK293T or HEK293T ATG2A/B KO cells after vehicle, AY9944 or rapamycin and chloroquine (Rap & CQN) treatment. Scale bar, 25 µm. (B) Quantified CD63 expression in HEK293T and HEK293T ATG2A/B KO cells (mean ± SEM; n = 8 images analyzed from 2 independent experiments). Two‐way ANOVA (treatment effect: F2,42 = 47.99, p < 0.0001; Autophagy effect: F1,42 = 5.422, p ≤ 0.0248). Sidak's multiple comparisons test (** p ≤ 0.005, **** p < 0.0001). (C) LC3B expression in HEK293T and HEK293T ATG2A/B KO cells (mean ± SEM; n = 8, 4 images analyzed from 2 independent experiments). Two‐way ANOVA (interaction effect: F2,42 = 6.126, p ≤ 0.0046; treatment effect: F2,42 = 5.995, p ≤ 0.0051; Autophagy effect: F1,42 = 12.65, p ≤ 0.0009). Sidak's multiple comparisons test (* p ≤ 0.05, ** p ≤ 0.005, *** p ≤ 0.001). (D) Immunoblot for LC3B in isolated sEVs upon cholesterol depletion. (E) CD63 and LC3B colocalization in HEK293T and HEK293T ATG2A/B KO cells (mean ± SEM; n = 8 images analyzed from 2 independent experiments). Two‐way ANOVA (interaction effect: F2,53 = 128.9, p < 0.0001; treatment effect: F2,53 = 157.0, p < 0.0001; autophagy effect: F1,53 = 453.1, p < 0.0001). Sidak's multiple comparisons test (**** p < 0.0001). (F) Quantified sEV secretion in HEK293T or HEK293T ATG2A/B KO cells following vehicle, AY9944, or Rap & CQN treatments (mean ± SEM; n = 3 biological replicates from 3 independent experiments). One‐way ANOVA (F2,6 = 56.17, p ≤ 0.0001), Dunnett's multiple comparisons test (** p ≤ 0.01, **** p < 0.0001). (G) sEV secretion in HEK293T or HEK293T ATG2A/B KO cells following vehicle or <t>D609</t> treatments (mean ± SEM; n = 3 biological replicates from 3 independent experiments). Unpaired t‐test (HEK293T: t4 = 9.395, *** p ≤ 0.0007; HEK293T ATG2A/B KO: t 4 = 4.069, ** p ≤ 0.0152).
    D609, supplied by MedChemExpress, used in various techniques. Bioz Stars score: 94/100, based on 17 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    d609 - by Bioz Stars, 2026-02
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    Images

    1) Product Images from "Cholesterol Deficiency Directs Autophagy‐Dependent Secretion of Extracellular Vesicles"

    Article Title: Cholesterol Deficiency Directs Autophagy‐Dependent Secretion of Extracellular Vesicles

    Journal: Journal of Extracellular Vesicles

    doi: 10.1002/jev2.70218

    Increased sEV secretion in cholesterol‐depleted conditions is due to autophagy‐driven signalling. (A) Representative images for CD63 and LC3B expression in HEK293T or HEK293T ATG2A/B KO cells after vehicle, AY9944 or rapamycin and chloroquine (Rap & CQN) treatment. Scale bar, 25 µm. (B) Quantified CD63 expression in HEK293T and HEK293T ATG2A/B KO cells (mean ± SEM; n = 8 images analyzed from 2 independent experiments). Two‐way ANOVA (treatment effect: F2,42 = 47.99, p < 0.0001; Autophagy effect: F1,42 = 5.422, p ≤ 0.0248). Sidak's multiple comparisons test (** p ≤ 0.005, **** p < 0.0001). (C) LC3B expression in HEK293T and HEK293T ATG2A/B KO cells (mean ± SEM; n = 8, 4 images analyzed from 2 independent experiments). Two‐way ANOVA (interaction effect: F2,42 = 6.126, p ≤ 0.0046; treatment effect: F2,42 = 5.995, p ≤ 0.0051; Autophagy effect: F1,42 = 12.65, p ≤ 0.0009). Sidak's multiple comparisons test (* p ≤ 0.05, ** p ≤ 0.005, *** p ≤ 0.001). (D) Immunoblot for LC3B in isolated sEVs upon cholesterol depletion. (E) CD63 and LC3B colocalization in HEK293T and HEK293T ATG2A/B KO cells (mean ± SEM; n = 8 images analyzed from 2 independent experiments). Two‐way ANOVA (interaction effect: F2,53 = 128.9, p < 0.0001; treatment effect: F2,53 = 157.0, p < 0.0001; autophagy effect: F1,53 = 453.1, p < 0.0001). Sidak's multiple comparisons test (**** p < 0.0001). (F) Quantified sEV secretion in HEK293T or HEK293T ATG2A/B KO cells following vehicle, AY9944, or Rap & CQN treatments (mean ± SEM; n = 3 biological replicates from 3 independent experiments). One‐way ANOVA (F2,6 = 56.17, p ≤ 0.0001), Dunnett's multiple comparisons test (** p ≤ 0.01, **** p < 0.0001). (G) sEV secretion in HEK293T or HEK293T ATG2A/B KO cells following vehicle or D609 treatments (mean ± SEM; n = 3 biological replicates from 3 independent experiments). Unpaired t‐test (HEK293T: t4 = 9.395, *** p ≤ 0.0007; HEK293T ATG2A/B KO: t 4 = 4.069, ** p ≤ 0.0152).
    Figure Legend Snippet: Increased sEV secretion in cholesterol‐depleted conditions is due to autophagy‐driven signalling. (A) Representative images for CD63 and LC3B expression in HEK293T or HEK293T ATG2A/B KO cells after vehicle, AY9944 or rapamycin and chloroquine (Rap & CQN) treatment. Scale bar, 25 µm. (B) Quantified CD63 expression in HEK293T and HEK293T ATG2A/B KO cells (mean ± SEM; n = 8 images analyzed from 2 independent experiments). Two‐way ANOVA (treatment effect: F2,42 = 47.99, p < 0.0001; Autophagy effect: F1,42 = 5.422, p ≤ 0.0248). Sidak's multiple comparisons test (** p ≤ 0.005, **** p < 0.0001). (C) LC3B expression in HEK293T and HEK293T ATG2A/B KO cells (mean ± SEM; n = 8, 4 images analyzed from 2 independent experiments). Two‐way ANOVA (interaction effect: F2,42 = 6.126, p ≤ 0.0046; treatment effect: F2,42 = 5.995, p ≤ 0.0051; Autophagy effect: F1,42 = 12.65, p ≤ 0.0009). Sidak's multiple comparisons test (* p ≤ 0.05, ** p ≤ 0.005, *** p ≤ 0.001). (D) Immunoblot for LC3B in isolated sEVs upon cholesterol depletion. (E) CD63 and LC3B colocalization in HEK293T and HEK293T ATG2A/B KO cells (mean ± SEM; n = 8 images analyzed from 2 independent experiments). Two‐way ANOVA (interaction effect: F2,53 = 128.9, p < 0.0001; treatment effect: F2,53 = 157.0, p < 0.0001; autophagy effect: F1,53 = 453.1, p < 0.0001). Sidak's multiple comparisons test (**** p < 0.0001). (F) Quantified sEV secretion in HEK293T or HEK293T ATG2A/B KO cells following vehicle, AY9944, or Rap & CQN treatments (mean ± SEM; n = 3 biological replicates from 3 independent experiments). One‐way ANOVA (F2,6 = 56.17, p ≤ 0.0001), Dunnett's multiple comparisons test (** p ≤ 0.01, **** p < 0.0001). (G) sEV secretion in HEK293T or HEK293T ATG2A/B KO cells following vehicle or D609 treatments (mean ± SEM; n = 3 biological replicates from 3 independent experiments). Unpaired t‐test (HEK293T: t4 = 9.395, *** p ≤ 0.0007; HEK293T ATG2A/B KO: t 4 = 4.069, ** p ≤ 0.0152).

    Techniques Used: Expressing, Western Blot, Isolation



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    Increased sEV secretion in cholesterol‐depleted conditions is due to autophagy‐driven signalling. (A) Representative images for CD63 and LC3B expression in HEK293T or HEK293T ATG2A/B KO cells after vehicle, AY9944 or rapamycin and chloroquine (Rap & CQN) treatment. Scale bar, 25 µm. (B) Quantified CD63 expression in HEK293T and HEK293T ATG2A/B KO cells (mean ± SEM; n = 8 images analyzed from 2 independent experiments). Two‐way ANOVA (treatment effect: F2,42 = 47.99, p < 0.0001; Autophagy effect: F1,42 = 5.422, p ≤ 0.0248). Sidak's multiple comparisons test (** p ≤ 0.005, **** p < 0.0001). (C) LC3B expression in HEK293T and HEK293T ATG2A/B KO cells (mean ± SEM; n = 8, 4 images analyzed from 2 independent experiments). Two‐way ANOVA (interaction effect: F2,42 = 6.126, p ≤ 0.0046; treatment effect: F2,42 = 5.995, p ≤ 0.0051; Autophagy effect: F1,42 = 12.65, p ≤ 0.0009). Sidak's multiple comparisons test (* p ≤ 0.05, ** p ≤ 0.005, *** p ≤ 0.001). (D) Immunoblot for LC3B in isolated sEVs upon cholesterol depletion. (E) CD63 and LC3B colocalization in HEK293T and HEK293T ATG2A/B KO cells (mean ± SEM; n = 8 images analyzed from 2 independent experiments). Two‐way ANOVA (interaction effect: F2,53 = 128.9, p < 0.0001; treatment effect: F2,53 = 157.0, p < 0.0001; autophagy effect: F1,53 = 453.1, p < 0.0001). Sidak's multiple comparisons test (**** p < 0.0001). (F) Quantified sEV secretion in HEK293T or HEK293T ATG2A/B KO cells following vehicle, AY9944, or Rap & CQN treatments (mean ± SEM; n = 3 biological replicates from 3 independent experiments). One‐way ANOVA (F2,6 = 56.17, p ≤ 0.0001), Dunnett's multiple comparisons test (** p ≤ 0.01, **** p < 0.0001). (G) sEV secretion in HEK293T or HEK293T ATG2A/B KO cells following vehicle or <t>D609</t> treatments (mean ± SEM; n = 3 biological replicates from 3 independent experiments). Unpaired t‐test (HEK293T: t4 = 9.395, *** p ≤ 0.0007; HEK293T ATG2A/B KO: t 4 = 4.069, ** p ≤ 0.0152).
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    90
    ChemScene llc tricyclodecan-9-yl-xanthogenate (d609
    Effects of <t>D609,</t> a PC-PLC/SMS inhibitor, on PC-PLC, PE-PLC, SMS, and CPES activities of SMS2. A , DG-generating activities of SMS2, which was purified in the presence of detergents, were analyzed in the presence of various concentrations of D609 (0, 0.25, 0.5, and 1 mM). Purified WT SMS2-TS (approximately 500 ng) was incubated in the presence of 16:0/18:1-PC alone, 16:0/18:1-PC and d18:1/18:0-Cer, 16:0/18:1-PE alone, or 16:0/18:1-PE and d18:1/18:0-Cer–containing detergent mixed micelles for 2 h at 37 °C. Produced 16:0/18:1-DG was quantified using LC–MS/MS. Values are presented as percentages of the activity of SMS2 in the absence of D609 (set at 100%). Values are presented as the mean ± SD (n = 6, technical replicates). ∗∗∗, p < 0.005 ( versus control (without D609)). One-way ANOVA with Dunnett’s post hoc test was used. B , SM-producing activities of SMS2, which was purified in the presence of detergents, in the presence of 16:0/18:1-PC and d18:1/18:0-Cer–containing detergent mixed micelles were analyzed in the presence of various concentrations of D609 (0, 0.25, 0.5, and 1 mM). Values are presented as percentages of the SMS activity of SMS2 in the absence of D609 (set to 100%). The values are presented as the mean ± SD (n = 4, technical replicates). ∗, p < 0.05; ∗∗, p < 0.01 ( versus control (without D609)). One-way ANOVA with Dunnett’s post hoc test was used. C , CPE-producing activities of SMS2, which was purified in the presence of detergents, in the presence of 16:0/18:1-PE and d18:1/18:0-Cer–containing detergent mixed micelles were analyzed in the presence of various concentrations of D609 (0, 0.25, 0.5, and 1 mM). Values are presented as percentages of the CPES activity of SMS2 in the absence of D609 (set at 100%). Values are presented as the mean ± SD (n = 6, technical replicates). ∗∗, p < 0.01; ∗∗∗, p < 0.005 ( versus control (without D609)). One-way ANOVA with Dunnett’s post hoc test was used. Cer, ceramide; CPE, ceramide phosphoethanolamine; CPES, CPE synthase; DG, diacylglycerol; PC, phosphatidylcholine; PE, phosphatidylethanolamine; PLC, phospholipase C; SM, sphingomyelin; SMS, sphingomyelin synthase; TS, Twin-Strep.
    Tricyclodecan 9 Yl Xanthogenate (D609, supplied by ChemScene llc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/tricyclodecan-9-yl-xanthogenate (d609/product/ChemScene llc
    Average 90 stars, based on 1 article reviews
    tricyclodecan-9-yl-xanthogenate (d609 - by Bioz Stars, 2026-02
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    tricyclodecan-9-yl-xanthogenate (d609)  (ChemScene llc)
    90
    ChemScene llc tricyclodecan-9-yl-xanthogenate (d609)
    Effects of <t>D609,</t> a PC-PLC/SMS inhibitor, on PC-PLC, PE-PLC, SMS, and CPES activities of SMS2. A , DG-generating activities of SMS2, which was purified in the presence of detergents, were analyzed in the presence of various concentrations of D609 (0, 0.25, 0.5, and 1 mM). Purified WT SMS2-TS (approximately 500 ng) was incubated in the presence of 16:0/18:1-PC alone, 16:0/18:1-PC and d18:1/18:0-Cer, 16:0/18:1-PE alone, or 16:0/18:1-PE and d18:1/18:0-Cer–containing detergent mixed micelles for 2 h at 37 °C. Produced 16:0/18:1-DG was quantified using LC–MS/MS. Values are presented as percentages of the activity of SMS2 in the absence of D609 (set at 100%). Values are presented as the mean ± SD (n = 6, technical replicates). ∗∗∗, p < 0.005 ( versus control (without D609)). One-way ANOVA with Dunnett’s post hoc test was used. B , SM-producing activities of SMS2, which was purified in the presence of detergents, in the presence of 16:0/18:1-PC and d18:1/18:0-Cer–containing detergent mixed micelles were analyzed in the presence of various concentrations of D609 (0, 0.25, 0.5, and 1 mM). Values are presented as percentages of the SMS activity of SMS2 in the absence of D609 (set to 100%). The values are presented as the mean ± SD (n = 4, technical replicates). ∗, p < 0.05; ∗∗, p < 0.01 ( versus control (without D609)). One-way ANOVA with Dunnett’s post hoc test was used. C , CPE-producing activities of SMS2, which was purified in the presence of detergents, in the presence of 16:0/18:1-PE and d18:1/18:0-Cer–containing detergent mixed micelles were analyzed in the presence of various concentrations of D609 (0, 0.25, 0.5, and 1 mM). Values are presented as percentages of the CPES activity of SMS2 in the absence of D609 (set at 100%). Values are presented as the mean ± SD (n = 6, technical replicates). ∗∗, p < 0.01; ∗∗∗, p < 0.005 ( versus control (without D609)). One-way ANOVA with Dunnett’s post hoc test was used. Cer, ceramide; CPE, ceramide phosphoethanolamine; CPES, CPE synthase; DG, diacylglycerol; PC, phosphatidylcholine; PE, phosphatidylethanolamine; PLC, phospholipase C; SM, sphingomyelin; SMS, sphingomyelin synthase; TS, Twin-Strep.
    Tricyclodecan 9 Yl Xanthogenate (D609), supplied by ChemScene llc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/tricyclodecan-9-yl-xanthogenate (d609)/product/ChemScene llc
    Average 90 stars, based on 1 article reviews
    tricyclodecan-9-yl-xanthogenate (d609) - by Bioz Stars, 2026-02
    90/100 stars
    		  Buy from Supplier

    Image Search Results


    Increased sEV secretion in cholesterol‐depleted conditions is due to autophagy‐driven signalling. (A) Representative images for CD63 and LC3B expression in HEK293T or HEK293T ATG2A/B KO cells after vehicle, AY9944 or rapamycin and chloroquine (Rap & CQN) treatment. Scale bar, 25 µm. (B) Quantified CD63 expression in HEK293T and HEK293T ATG2A/B KO cells (mean ± SEM; n = 8 images analyzed from 2 independent experiments). Two‐way ANOVA (treatment effect: F2,42 = 47.99, p < 0.0001; Autophagy effect: F1,42 = 5.422, p ≤ 0.0248). Sidak's multiple comparisons test (** p ≤ 0.005, **** p < 0.0001). (C) LC3B expression in HEK293T and HEK293T ATG2A/B KO cells (mean ± SEM; n = 8, 4 images analyzed from 2 independent experiments). Two‐way ANOVA (interaction effect: F2,42 = 6.126, p ≤ 0.0046; treatment effect: F2,42 = 5.995, p ≤ 0.0051; Autophagy effect: F1,42 = 12.65, p ≤ 0.0009). Sidak's multiple comparisons test (* p ≤ 0.05, ** p ≤ 0.005, *** p ≤ 0.001). (D) Immunoblot for LC3B in isolated sEVs upon cholesterol depletion. (E) CD63 and LC3B colocalization in HEK293T and HEK293T ATG2A/B KO cells (mean ± SEM; n = 8 images analyzed from 2 independent experiments). Two‐way ANOVA (interaction effect: F2,53 = 128.9, p < 0.0001; treatment effect: F2,53 = 157.0, p < 0.0001; autophagy effect: F1,53 = 453.1, p < 0.0001). Sidak's multiple comparisons test (**** p < 0.0001). (F) Quantified sEV secretion in HEK293T or HEK293T ATG2A/B KO cells following vehicle, AY9944, or Rap & CQN treatments (mean ± SEM; n = 3 biological replicates from 3 independent experiments). One‐way ANOVA (F2,6 = 56.17, p ≤ 0.0001), Dunnett's multiple comparisons test (** p ≤ 0.01, **** p < 0.0001). (G) sEV secretion in HEK293T or HEK293T ATG2A/B KO cells following vehicle or D609 treatments (mean ± SEM; n = 3 biological replicates from 3 independent experiments). Unpaired t‐test (HEK293T: t4 = 9.395, *** p ≤ 0.0007; HEK293T ATG2A/B KO: t 4 = 4.069, ** p ≤ 0.0152).

    Journal: Journal of Extracellular Vesicles

    Article Title: Cholesterol Deficiency Directs Autophagy‐Dependent Secretion of Extracellular Vesicles

    doi: 10.1002/jev2.70218

    Figure Lengend Snippet: Increased sEV secretion in cholesterol‐depleted conditions is due to autophagy‐driven signalling. (A) Representative images for CD63 and LC3B expression in HEK293T or HEK293T ATG2A/B KO cells after vehicle, AY9944 or rapamycin and chloroquine (Rap & CQN) treatment. Scale bar, 25 µm. (B) Quantified CD63 expression in HEK293T and HEK293T ATG2A/B KO cells (mean ± SEM; n = 8 images analyzed from 2 independent experiments). Two‐way ANOVA (treatment effect: F2,42 = 47.99, p < 0.0001; Autophagy effect: F1,42 = 5.422, p ≤ 0.0248). Sidak's multiple comparisons test (** p ≤ 0.005, **** p < 0.0001). (C) LC3B expression in HEK293T and HEK293T ATG2A/B KO cells (mean ± SEM; n = 8, 4 images analyzed from 2 independent experiments). Two‐way ANOVA (interaction effect: F2,42 = 6.126, p ≤ 0.0046; treatment effect: F2,42 = 5.995, p ≤ 0.0051; Autophagy effect: F1,42 = 12.65, p ≤ 0.0009). Sidak's multiple comparisons test (* p ≤ 0.05, ** p ≤ 0.005, *** p ≤ 0.001). (D) Immunoblot for LC3B in isolated sEVs upon cholesterol depletion. (E) CD63 and LC3B colocalization in HEK293T and HEK293T ATG2A/B KO cells (mean ± SEM; n = 8 images analyzed from 2 independent experiments). Two‐way ANOVA (interaction effect: F2,53 = 128.9, p < 0.0001; treatment effect: F2,53 = 157.0, p < 0.0001; autophagy effect: F1,53 = 453.1, p < 0.0001). Sidak's multiple comparisons test (**** p < 0.0001). (F) Quantified sEV secretion in HEK293T or HEK293T ATG2A/B KO cells following vehicle, AY9944, or Rap & CQN treatments (mean ± SEM; n = 3 biological replicates from 3 independent experiments). One‐way ANOVA (F2,6 = 56.17, p ≤ 0.0001), Dunnett's multiple comparisons test (** p ≤ 0.01, **** p < 0.0001). (G) sEV secretion in HEK293T or HEK293T ATG2A/B KO cells following vehicle or D609 treatments (mean ± SEM; n = 3 biological replicates from 3 independent experiments). Unpaired t‐test (HEK293T: t4 = 9.395, *** p ≤ 0.0007; HEK293T ATG2A/B KO: t 4 = 4.069, ** p ≤ 0.0152).

    Article Snippet: For FBS rescue experiments, HEK293T cells were treated with vehicle, simvastatin or AY9944 for 24 h. Media was then replaced with 10% EV‐depleted FBS for 24 or 48 h. Other pharmacological agents utilized in these studies include rapamycin (1 μM, Selleck Chemicals; mTOR inhibitor), chloroquine (10 μM, Cayman Chemical; inhibits autophagosome‐lysosome fusion), D609 (100 μM, MedChemExpress; phosphatidyl choline phospholipase C inhibitor), and Q‐VD‐Oph (10 μM, Selleck Chemicals; pan‐caspase inhibitor).

    Techniques: Expressing, Western Blot, Isolation

    Effects of D609, a PC-PLC/SMS inhibitor, on PC-PLC, PE-PLC, SMS, and CPES activities of SMS2. A , DG-generating activities of SMS2, which was purified in the presence of detergents, were analyzed in the presence of various concentrations of D609 (0, 0.25, 0.5, and 1 mM). Purified WT SMS2-TS (approximately 500 ng) was incubated in the presence of 16:0/18:1-PC alone, 16:0/18:1-PC and d18:1/18:0-Cer, 16:0/18:1-PE alone, or 16:0/18:1-PE and d18:1/18:0-Cer–containing detergent mixed micelles for 2 h at 37 °C. Produced 16:0/18:1-DG was quantified using LC–MS/MS. Values are presented as percentages of the activity of SMS2 in the absence of D609 (set at 100%). Values are presented as the mean ± SD (n = 6, technical replicates). ∗∗∗, p < 0.005 ( versus control (without D609)). One-way ANOVA with Dunnett’s post hoc test was used. B , SM-producing activities of SMS2, which was purified in the presence of detergents, in the presence of 16:0/18:1-PC and d18:1/18:0-Cer–containing detergent mixed micelles were analyzed in the presence of various concentrations of D609 (0, 0.25, 0.5, and 1 mM). Values are presented as percentages of the SMS activity of SMS2 in the absence of D609 (set to 100%). The values are presented as the mean ± SD (n = 4, technical replicates). ∗, p < 0.05; ∗∗, p < 0.01 ( versus control (without D609)). One-way ANOVA with Dunnett’s post hoc test was used. C , CPE-producing activities of SMS2, which was purified in the presence of detergents, in the presence of 16:0/18:1-PE and d18:1/18:0-Cer–containing detergent mixed micelles were analyzed in the presence of various concentrations of D609 (0, 0.25, 0.5, and 1 mM). Values are presented as percentages of the CPES activity of SMS2 in the absence of D609 (set at 100%). Values are presented as the mean ± SD (n = 6, technical replicates). ∗∗, p < 0.01; ∗∗∗, p < 0.005 ( versus control (without D609)). One-way ANOVA with Dunnett’s post hoc test was used. Cer, ceramide; CPE, ceramide phosphoethanolamine; CPES, CPE synthase; DG, diacylglycerol; PC, phosphatidylcholine; PE, phosphatidylethanolamine; PLC, phospholipase C; SM, sphingomyelin; SMS, sphingomyelin synthase; TS, Twin-Strep.

    Journal: The Journal of Biological Chemistry

    Article Title: Multiple activities of sphingomyelin synthase 2 generate saturated fatty acid– and/or monounsaturated fatty acid–containing diacylglycerol

    doi: 10.1016/j.jbc.2024.107960

    Figure Lengend Snippet: Effects of D609, a PC-PLC/SMS inhibitor, on PC-PLC, PE-PLC, SMS, and CPES activities of SMS2. A , DG-generating activities of SMS2, which was purified in the presence of detergents, were analyzed in the presence of various concentrations of D609 (0, 0.25, 0.5, and 1 mM). Purified WT SMS2-TS (approximately 500 ng) was incubated in the presence of 16:0/18:1-PC alone, 16:0/18:1-PC and d18:1/18:0-Cer, 16:0/18:1-PE alone, or 16:0/18:1-PE and d18:1/18:0-Cer–containing detergent mixed micelles for 2 h at 37 °C. Produced 16:0/18:1-DG was quantified using LC–MS/MS. Values are presented as percentages of the activity of SMS2 in the absence of D609 (set at 100%). Values are presented as the mean ± SD (n = 6, technical replicates). ∗∗∗, p < 0.005 ( versus control (without D609)). One-way ANOVA with Dunnett’s post hoc test was used. B , SM-producing activities of SMS2, which was purified in the presence of detergents, in the presence of 16:0/18:1-PC and d18:1/18:0-Cer–containing detergent mixed micelles were analyzed in the presence of various concentrations of D609 (0, 0.25, 0.5, and 1 mM). Values are presented as percentages of the SMS activity of SMS2 in the absence of D609 (set to 100%). The values are presented as the mean ± SD (n = 4, technical replicates). ∗, p < 0.05; ∗∗, p < 0.01 ( versus control (without D609)). One-way ANOVA with Dunnett’s post hoc test was used. C , CPE-producing activities of SMS2, which was purified in the presence of detergents, in the presence of 16:0/18:1-PE and d18:1/18:0-Cer–containing detergent mixed micelles were analyzed in the presence of various concentrations of D609 (0, 0.25, 0.5, and 1 mM). Values are presented as percentages of the CPES activity of SMS2 in the absence of D609 (set at 100%). Values are presented as the mean ± SD (n = 6, technical replicates). ∗∗, p < 0.01; ∗∗∗, p < 0.005 ( versus control (without D609)). One-way ANOVA with Dunnett’s post hoc test was used. Cer, ceramide; CPE, ceramide phosphoethanolamine; CPES, CPE synthase; DG, diacylglycerol; PC, phosphatidylcholine; PE, phosphatidylethanolamine; PLC, phospholipase C; SM, sphingomyelin; SMS, sphingomyelin synthase; TS, Twin-Strep.

    Article Snippet: Tricyclodecan-9-yl-xanthogenate (D609) was obtained from Chemscene.

    Techniques: Purification, Incubation, Produced, Liquid Chromatography with Mass Spectroscopy, Activity Assay, Control

    Comparison of SMS2 (our findings and previous reports), SMS1, and SMSr

    Journal: The Journal of Biological Chemistry

    Article Title: Multiple activities of sphingomyelin synthase 2 generate saturated fatty acid– and/or monounsaturated fatty acid–containing diacylglycerol

    doi: 10.1016/j.jbc.2024.107960

    Figure Lengend Snippet: Comparison of SMS2 (our findings and previous reports), SMS1, and SMSr

    Article Snippet: Tricyclodecan-9-yl-xanthogenate (D609) was obtained from Chemscene.

    Techniques: Comparison, Membrane, Expressing, Activity Assay, In Vitro, Inhibition, Mutagenesis

    Comparison of bacterial, plant, and mammalian PLCs

    Journal: The Journal of Biological Chemistry

    Article Title: Multiple activities of sphingomyelin synthase 2 generate saturated fatty acid– and/or monounsaturated fatty acid–containing diacylglycerol

    doi: 10.1016/j.jbc.2024.107960

    Figure Lengend Snippet: Comparison of bacterial, plant, and mammalian PLCs

    Article Snippet: Tricyclodecan-9-yl-xanthogenate (D609) was obtained from Chemscene.

    Techniques: Comparison, Activity Assay, Membrane, Inhibition