Review



elovl6 inhibitor elovl6  (MedChemExpress)


Bioz Verified Symbol MedChemExpress is a verified supplier
Bioz Manufacturer Symbol MedChemExpress manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
    • 94
      Buy from Supplier

    Structured Review

    MedChemExpress elovl6 inhibitor elovl6
    OA induces PLD activation in HEK293T and MDA-MB-231 cells. A, B: Representative confocal microscopy images of the PA sensor (RFP-PASS in HEK293T cells and GFP-PASS in MDA-MB-231 cells) in response to FA treatments. Cells were transiently transfected with RFP/GFP-PASS. After 48 h incubation, cells were starved with serum-free medium for 3 h with 1 μM SCD1 inhibitor A939572 and 1 μM <t>ELOVL6</t> inhibitor ELOVL6-IN-4 to prevent FA desaturation and elongation (for lipid raft disruption, cells were preincubated with 100 μM MβCD in serum-free medium for 3 h). Cells were then incubated with 100 μM BSA-conjugated FAs (BSA for Ctrl) in the continued presence of these inhibitors. Scale bars in the first micrograph apply to all images in series A or B. C: Quantification of RFP/GFP-PASS fluorescence on the plasma membrane (PM) relative to the cytoplasm. The average (± standard deviation) ratio was calculated from three independent experiments (HEK293T, n ≥ 60; MDA-MB-231, n ≥ 33). Statistical significance was assessed using one-way ANOVA followed by Dunnett’s multiple comparisons test, with significance thresholds defined as follows: ns, not significant, P ≥ 0.05; ∗, P < 0.05; ∗∗∗∗, P < 0.0001.
    Elovl6 Inhibitor Elovl6, supplied by MedChemExpress, used in various techniques. Bioz Stars score: 94/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/elovl6 inhibitor elovl6/product/MedChemExpress
    Average 94 stars, based on 2 article reviews
    elovl6 inhibitor elovl6 - by Bioz Stars, 2026-02
    94/100 stars

    Images

    1) Product Images from "Oleate activates PLD2 lipase and GEF activity by modulating membrane microdomain dynamics via S-acylation"

    Article Title: Oleate activates PLD2 lipase and GEF activity by modulating membrane microdomain dynamics via S-acylation

    Journal: Journal of Lipid Research

    doi: 10.1016/j.jlr.2025.100939

    OA induces PLD activation in HEK293T and MDA-MB-231 cells. A, B: Representative confocal microscopy images of the PA sensor (RFP-PASS in HEK293T cells and GFP-PASS in MDA-MB-231 cells) in response to FA treatments. Cells were transiently transfected with RFP/GFP-PASS. After 48 h incubation, cells were starved with serum-free medium for 3 h with 1 μM SCD1 inhibitor A939572 and 1 μM ELOVL6 inhibitor ELOVL6-IN-4 to prevent FA desaturation and elongation (for lipid raft disruption, cells were preincubated with 100 μM MβCD in serum-free medium for 3 h). Cells were then incubated with 100 μM BSA-conjugated FAs (BSA for Ctrl) in the continued presence of these inhibitors. Scale bars in the first micrograph apply to all images in series A or B. C: Quantification of RFP/GFP-PASS fluorescence on the plasma membrane (PM) relative to the cytoplasm. The average (± standard deviation) ratio was calculated from three independent experiments (HEK293T, n ≥ 60; MDA-MB-231, n ≥ 33). Statistical significance was assessed using one-way ANOVA followed by Dunnett’s multiple comparisons test, with significance thresholds defined as follows: ns, not significant, P ≥ 0.05; ∗, P < 0.05; ∗∗∗∗, P < 0.0001.
    Figure Legend Snippet: OA induces PLD activation in HEK293T and MDA-MB-231 cells. A, B: Representative confocal microscopy images of the PA sensor (RFP-PASS in HEK293T cells and GFP-PASS in MDA-MB-231 cells) in response to FA treatments. Cells were transiently transfected with RFP/GFP-PASS. After 48 h incubation, cells were starved with serum-free medium for 3 h with 1 μM SCD1 inhibitor A939572 and 1 μM ELOVL6 inhibitor ELOVL6-IN-4 to prevent FA desaturation and elongation (for lipid raft disruption, cells were preincubated with 100 μM MβCD in serum-free medium for 3 h). Cells were then incubated with 100 μM BSA-conjugated FAs (BSA for Ctrl) in the continued presence of these inhibitors. Scale bars in the first micrograph apply to all images in series A or B. C: Quantification of RFP/GFP-PASS fluorescence on the plasma membrane (PM) relative to the cytoplasm. The average (± standard deviation) ratio was calculated from three independent experiments (HEK293T, n ≥ 60; MDA-MB-231, n ≥ 33). Statistical significance was assessed using one-way ANOVA followed by Dunnett’s multiple comparisons test, with significance thresholds defined as follows: ns, not significant, P ≥ 0.05; ∗, P < 0.05; ∗∗∗∗, P < 0.0001.

    Techniques Used: Activation Assay, Confocal Microscopy, Transfection, Incubation, Disruption, Fluorescence, Clinical Proteomics, Membrane, Standard Deviation

    OA disrupts the colocalization of PLD2 with lipid rafts. A: Representative confocal microscopy images showing the colocalization of PLD2 (conjugated with Alexa Fluor 647 secondary antibody, in red) and Caveolin-1 (conjugated with Alexa Fluor 488 secondary antibody, in green) in HEK293T cells and MDA-MB-231 cells treated 3 h with BSA (Ctrl) or 100 μM OA. Scale bars in the first micrograph apply to all images in the series. The zoomed-in channels correspond to the regions highlighted by the yellow rectangles in the merged image. B, C: Quantification of caveolin-1 and PLD2 colocalization (B) and distribution variability (C). Colocalization was assessed using Pearson's correlation coefficient. Distribution heterogeneity was measured by coefficient of variation (CV). D, E: Western blot analysis of lipid raft fractions isolated by sucrose density gradient ultracentrifugation from HEK293T cells expressing EGFP-PLD2 (D) and MDA-MB-231 cells (E). Fractions 3–5 are enriched in lipid rafts, as indicated by the predominant presence of Caveolin-1. Bar charts show the percentage of PLD2 localized in lipid raft fractions under BSA (Ctrl), C18:0, C18:1, C16:0, and C16:1 treatments. Cells were pretreated with 1 μM SCD1 inhibitor A939572 and 1 μM ELOVL6 inhibitor ELOVL6-IN-4 in serum-free DMEM for 3 h to prevent FA desaturation and elongation. 10 μM palmostatin B (Palm B) was used to preserve S-acylated residues in cell lysis buffer. The proportion of PLD2 in raft fractions was calculated as the ratio of the sum of PLD2 in raft fractions (fractions 3–5) to the sum of PLD2 in all 11 fractions. Data are presented as mean ± SEM (n = 3 independent experiments). Statistical significance was assessed using unpaired two-tailed t-tests (B–C) or one-way ANOVA followed by Dunnett’s multiple comparisons test (D–E), with significance thresholds defined as follows: ns, not significant, P ≥ 0.05; ∗, P < 0.05; ∗∗, P < 0.01; ∗∗∗, P < 0.001.
    Figure Legend Snippet: OA disrupts the colocalization of PLD2 with lipid rafts. A: Representative confocal microscopy images showing the colocalization of PLD2 (conjugated with Alexa Fluor 647 secondary antibody, in red) and Caveolin-1 (conjugated with Alexa Fluor 488 secondary antibody, in green) in HEK293T cells and MDA-MB-231 cells treated 3 h with BSA (Ctrl) or 100 μM OA. Scale bars in the first micrograph apply to all images in the series. The zoomed-in channels correspond to the regions highlighted by the yellow rectangles in the merged image. B, C: Quantification of caveolin-1 and PLD2 colocalization (B) and distribution variability (C). Colocalization was assessed using Pearson's correlation coefficient. Distribution heterogeneity was measured by coefficient of variation (CV). D, E: Western blot analysis of lipid raft fractions isolated by sucrose density gradient ultracentrifugation from HEK293T cells expressing EGFP-PLD2 (D) and MDA-MB-231 cells (E). Fractions 3–5 are enriched in lipid rafts, as indicated by the predominant presence of Caveolin-1. Bar charts show the percentage of PLD2 localized in lipid raft fractions under BSA (Ctrl), C18:0, C18:1, C16:0, and C16:1 treatments. Cells were pretreated with 1 μM SCD1 inhibitor A939572 and 1 μM ELOVL6 inhibitor ELOVL6-IN-4 in serum-free DMEM for 3 h to prevent FA desaturation and elongation. 10 μM palmostatin B (Palm B) was used to preserve S-acylated residues in cell lysis buffer. The proportion of PLD2 in raft fractions was calculated as the ratio of the sum of PLD2 in raft fractions (fractions 3–5) to the sum of PLD2 in all 11 fractions. Data are presented as mean ± SEM (n = 3 independent experiments). Statistical significance was assessed using unpaired two-tailed t-tests (B–C) or one-way ANOVA followed by Dunnett’s multiple comparisons test (D–E), with significance thresholds defined as follows: ns, not significant, P ≥ 0.05; ∗, P < 0.05; ∗∗, P < 0.01; ∗∗∗, P < 0.001.

    Techniques Used: Confocal Microscopy, Western Blot, Isolation, Expressing, Lysis, Two Tailed Test

    The impact of PLD2 S-acylation site mutation on its targeting to lipid rafts. A: Western blot analysis of lipid raft fractions isolated by sucrose density gradient ultracentrifugation from HEK293T cells expressing EGFP-PLD2 (WT, C223A, C224A, and C223AC224A). Cells were starved with serum-free medium and preincubated with 1 μM SCD1 inhibitor A939572 and 1 μM ELOVL6 inhibitor ELOVL6-IN-4 for 3 h to prevent FA desaturation and elongation. Cells were then treated with BSA (Ctrl groups) or FAs (100 μM OA for WT, C223A, C224A; 100 μM palmitate for C223AC224A) for 3 h. Fractions 3–5 pinpoint lipid raft-enriched fractions, as indicated by the presence of Caveolin-1. B: Quantification of the proportion of PLD2 in raft fractions relative to the total PLD2 across all fractions. C: Confocal microscopy images of HEK293T cells expressing EGFP-PLD2-WT and -C223AC224A. Cells were then stained with anti-PIP 2 antibody (conjugated with Alexa Fluor 647 secondary antibody) to assess colocalization with EGFP-PLD2. The scale bar in the first micrograph applies to all images in the series of panel. The zoomed-in channels correspond to the regions highlighted by the yellow rectangles in the merged image. Colocalization index was assessed using Pearson's correlation coefficient. Data are presented as mean ± SEM (n = 3 independent experiments). Statistical significances were analyzed using one-way ANOVA followed by Dunnett’s multiple comparisons test to WT in Ctrl group for (B). Unpaired two-tailed Student's t-tests were used in FA treated group to compare the FA treated group and the corresponding BSA-treated control for (B) and WT versus C223AC224A for (C). Significance thresholds were defined as: ns, not significant, P ≥ 0.05; ∗∗, P < 0.01; ∗∗∗∗, P < 0.0001.
    Figure Legend Snippet: The impact of PLD2 S-acylation site mutation on its targeting to lipid rafts. A: Western blot analysis of lipid raft fractions isolated by sucrose density gradient ultracentrifugation from HEK293T cells expressing EGFP-PLD2 (WT, C223A, C224A, and C223AC224A). Cells were starved with serum-free medium and preincubated with 1 μM SCD1 inhibitor A939572 and 1 μM ELOVL6 inhibitor ELOVL6-IN-4 for 3 h to prevent FA desaturation and elongation. Cells were then treated with BSA (Ctrl groups) or FAs (100 μM OA for WT, C223A, C224A; 100 μM palmitate for C223AC224A) for 3 h. Fractions 3–5 pinpoint lipid raft-enriched fractions, as indicated by the presence of Caveolin-1. B: Quantification of the proportion of PLD2 in raft fractions relative to the total PLD2 across all fractions. C: Confocal microscopy images of HEK293T cells expressing EGFP-PLD2-WT and -C223AC224A. Cells were then stained with anti-PIP 2 antibody (conjugated with Alexa Fluor 647 secondary antibody) to assess colocalization with EGFP-PLD2. The scale bar in the first micrograph applies to all images in the series of panel. The zoomed-in channels correspond to the regions highlighted by the yellow rectangles in the merged image. Colocalization index was assessed using Pearson's correlation coefficient. Data are presented as mean ± SEM (n = 3 independent experiments). Statistical significances were analyzed using one-way ANOVA followed by Dunnett’s multiple comparisons test to WT in Ctrl group for (B). Unpaired two-tailed Student's t-tests were used in FA treated group to compare the FA treated group and the corresponding BSA-treated control for (B) and WT versus C223AC224A for (C). Significance thresholds were defined as: ns, not significant, P ≥ 0.05; ∗∗, P < 0.01; ∗∗∗∗, P < 0.0001.

    Techniques Used: Mutagenesis, Western Blot, Isolation, Expressing, Confocal Microscopy, Staining, Two Tailed Test, Control



    Similar Products

    elovl6 inhibitor elovl6  (MedChemExpress)
    94
    MedChemExpress elovl6 inhibitor elovl6
    OA induces PLD activation in HEK293T and MDA-MB-231 cells. A, B: Representative confocal microscopy images of the PA sensor (RFP-PASS in HEK293T cells and GFP-PASS in MDA-MB-231 cells) in response to FA treatments. Cells were transiently transfected with RFP/GFP-PASS. After 48 h incubation, cells were starved with serum-free medium for 3 h with 1 μM SCD1 inhibitor A939572 and 1 μM <t>ELOVL6</t> inhibitor ELOVL6-IN-4 to prevent FA desaturation and elongation (for lipid raft disruption, cells were preincubated with 100 μM MβCD in serum-free medium for 3 h). Cells were then incubated with 100 μM BSA-conjugated FAs (BSA for Ctrl) in the continued presence of these inhibitors. Scale bars in the first micrograph apply to all images in series A or B. C: Quantification of RFP/GFP-PASS fluorescence on the plasma membrane (PM) relative to the cytoplasm. The average (± standard deviation) ratio was calculated from three independent experiments (HEK293T, n ≥ 60; MDA-MB-231, n ≥ 33). Statistical significance was assessed using one-way ANOVA followed by Dunnett’s multiple comparisons test, with significance thresholds defined as follows: ns, not significant, P ≥ 0.05; ∗, P < 0.05; ∗∗∗∗, P < 0.0001.
    Elovl6 Inhibitor Elovl6, supplied by MedChemExpress, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/elovl6 inhibitor elovl6/product/MedChemExpress
    Average 94 stars, based on 1 article reviews
    elovl6 inhibitor elovl6 - by Bioz Stars, 2026-02
    94/100 stars
    		  Buy from Supplier
    elovl6-in-4  (MedChemExpress)
    94
    MedChemExpress elovl6-in-4
    OA induces PLD activation in HEK293T and MDA-MB-231 cells. A, B: Representative confocal microscopy images of the PA sensor (RFP-PASS in HEK293T cells and GFP-PASS in MDA-MB-231 cells) in response to FA treatments. Cells were transiently transfected with RFP/GFP-PASS. After 48 h incubation, cells were starved with serum-free medium for 3 h with 1 μM SCD1 inhibitor A939572 and 1 μM <t>ELOVL6</t> inhibitor ELOVL6-IN-4 to prevent FA desaturation and elongation (for lipid raft disruption, cells were preincubated with 100 μM MβCD in serum-free medium for 3 h). Cells were then incubated with 100 μM BSA-conjugated FAs (BSA for Ctrl) in the continued presence of these inhibitors. Scale bars in the first micrograph apply to all images in series A or B. C: Quantification of RFP/GFP-PASS fluorescence on the plasma membrane (PM) relative to the cytoplasm. The average (± standard deviation) ratio was calculated from three independent experiments (HEK293T, n ≥ 60; MDA-MB-231, n ≥ 33). Statistical significance was assessed using one-way ANOVA followed by Dunnett’s multiple comparisons test, with significance thresholds defined as follows: ns, not significant, P ≥ 0.05; ∗, P < 0.05; ∗∗∗∗, P < 0.0001.
    Elovl6 In 4, supplied by MedChemExpress, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/elovl6-in-4/product/MedChemExpress
    Average 94 stars, based on 1 article reviews
    elovl6-in-4 - by Bioz Stars, 2026-02
    94/100 stars
    		  Buy from Supplier

    Image Search Results


    OA induces PLD activation in HEK293T and MDA-MB-231 cells. A, B: Representative confocal microscopy images of the PA sensor (RFP-PASS in HEK293T cells and GFP-PASS in MDA-MB-231 cells) in response to FA treatments. Cells were transiently transfected with RFP/GFP-PASS. After 48 h incubation, cells were starved with serum-free medium for 3 h with 1 μM SCD1 inhibitor A939572 and 1 μM ELOVL6 inhibitor ELOVL6-IN-4 to prevent FA desaturation and elongation (for lipid raft disruption, cells were preincubated with 100 μM MβCD in serum-free medium for 3 h). Cells were then incubated with 100 μM BSA-conjugated FAs (BSA for Ctrl) in the continued presence of these inhibitors. Scale bars in the first micrograph apply to all images in series A or B. C: Quantification of RFP/GFP-PASS fluorescence on the plasma membrane (PM) relative to the cytoplasm. The average (± standard deviation) ratio was calculated from three independent experiments (HEK293T, n ≥ 60; MDA-MB-231, n ≥ 33). Statistical significance was assessed using one-way ANOVA followed by Dunnett’s multiple comparisons test, with significance thresholds defined as follows: ns, not significant, P ≥ 0.05; ∗, P < 0.05; ∗∗∗∗, P < 0.0001.

    Journal: Journal of Lipid Research

    Article Title: Oleate activates PLD2 lipase and GEF activity by modulating membrane microdomain dynamics via S-acylation

    doi: 10.1016/j.jlr.2025.100939

    Figure Lengend Snippet: OA induces PLD activation in HEK293T and MDA-MB-231 cells. A, B: Representative confocal microscopy images of the PA sensor (RFP-PASS in HEK293T cells and GFP-PASS in MDA-MB-231 cells) in response to FA treatments. Cells were transiently transfected with RFP/GFP-PASS. After 48 h incubation, cells were starved with serum-free medium for 3 h with 1 μM SCD1 inhibitor A939572 and 1 μM ELOVL6 inhibitor ELOVL6-IN-4 to prevent FA desaturation and elongation (for lipid raft disruption, cells were preincubated with 100 μM MβCD in serum-free medium for 3 h). Cells were then incubated with 100 μM BSA-conjugated FAs (BSA for Ctrl) in the continued presence of these inhibitors. Scale bars in the first micrograph apply to all images in series A or B. C: Quantification of RFP/GFP-PASS fluorescence on the plasma membrane (PM) relative to the cytoplasm. The average (± standard deviation) ratio was calculated from three independent experiments (HEK293T, n ≥ 60; MDA-MB-231, n ≥ 33). Statistical significance was assessed using one-way ANOVA followed by Dunnett’s multiple comparisons test, with significance thresholds defined as follows: ns, not significant, P ≥ 0.05; ∗, P < 0.05; ∗∗∗∗, P < 0.0001.

    Article Snippet: Additional reagents and kits were obtained as follows: Cdc42 Activation Assay Kit (#8819, Cell Signaling Technology), Bradford protein assay (Bio-Rad, #5000006), Phalloidin–TRITC (Sigma, #P1951), tris(2-carboxyethyl)phosphine hydrochloride (TCEP; Sigma, #C4706), methoxypolyethylene glycol maleimide (mPEG–Mal; Sigma, #63187), N-ethylmaleimide (NEM; Sigma, #E3876), protease and phosphatase inhibitors (Sigma, #P8340; #P0044), ELOVL6 inhibitor ELOVL6-IN-4 (MedChemExpress, #HY-152947), and SCD1 inhibitor A939572 (Biofine, #37062).

    Techniques: Activation Assay, Confocal Microscopy, Transfection, Incubation, Disruption, Fluorescence, Clinical Proteomics, Membrane, Standard Deviation

    OA disrupts the colocalization of PLD2 with lipid rafts. A: Representative confocal microscopy images showing the colocalization of PLD2 (conjugated with Alexa Fluor 647 secondary antibody, in red) and Caveolin-1 (conjugated with Alexa Fluor 488 secondary antibody, in green) in HEK293T cells and MDA-MB-231 cells treated 3 h with BSA (Ctrl) or 100 μM OA. Scale bars in the first micrograph apply to all images in the series. The zoomed-in channels correspond to the regions highlighted by the yellow rectangles in the merged image. B, C: Quantification of caveolin-1 and PLD2 colocalization (B) and distribution variability (C). Colocalization was assessed using Pearson's correlation coefficient. Distribution heterogeneity was measured by coefficient of variation (CV). D, E: Western blot analysis of lipid raft fractions isolated by sucrose density gradient ultracentrifugation from HEK293T cells expressing EGFP-PLD2 (D) and MDA-MB-231 cells (E). Fractions 3–5 are enriched in lipid rafts, as indicated by the predominant presence of Caveolin-1. Bar charts show the percentage of PLD2 localized in lipid raft fractions under BSA (Ctrl), C18:0, C18:1, C16:0, and C16:1 treatments. Cells were pretreated with 1 μM SCD1 inhibitor A939572 and 1 μM ELOVL6 inhibitor ELOVL6-IN-4 in serum-free DMEM for 3 h to prevent FA desaturation and elongation. 10 μM palmostatin B (Palm B) was used to preserve S-acylated residues in cell lysis buffer. The proportion of PLD2 in raft fractions was calculated as the ratio of the sum of PLD2 in raft fractions (fractions 3–5) to the sum of PLD2 in all 11 fractions. Data are presented as mean ± SEM (n = 3 independent experiments). Statistical significance was assessed using unpaired two-tailed t-tests (B–C) or one-way ANOVA followed by Dunnett’s multiple comparisons test (D–E), with significance thresholds defined as follows: ns, not significant, P ≥ 0.05; ∗, P < 0.05; ∗∗, P < 0.01; ∗∗∗, P < 0.001.

    Journal: Journal of Lipid Research

    Article Title: Oleate activates PLD2 lipase and GEF activity by modulating membrane microdomain dynamics via S-acylation

    doi: 10.1016/j.jlr.2025.100939

    Figure Lengend Snippet: OA disrupts the colocalization of PLD2 with lipid rafts. A: Representative confocal microscopy images showing the colocalization of PLD2 (conjugated with Alexa Fluor 647 secondary antibody, in red) and Caveolin-1 (conjugated with Alexa Fluor 488 secondary antibody, in green) in HEK293T cells and MDA-MB-231 cells treated 3 h with BSA (Ctrl) or 100 μM OA. Scale bars in the first micrograph apply to all images in the series. The zoomed-in channels correspond to the regions highlighted by the yellow rectangles in the merged image. B, C: Quantification of caveolin-1 and PLD2 colocalization (B) and distribution variability (C). Colocalization was assessed using Pearson's correlation coefficient. Distribution heterogeneity was measured by coefficient of variation (CV). D, E: Western blot analysis of lipid raft fractions isolated by sucrose density gradient ultracentrifugation from HEK293T cells expressing EGFP-PLD2 (D) and MDA-MB-231 cells (E). Fractions 3–5 are enriched in lipid rafts, as indicated by the predominant presence of Caveolin-1. Bar charts show the percentage of PLD2 localized in lipid raft fractions under BSA (Ctrl), C18:0, C18:1, C16:0, and C16:1 treatments. Cells were pretreated with 1 μM SCD1 inhibitor A939572 and 1 μM ELOVL6 inhibitor ELOVL6-IN-4 in serum-free DMEM for 3 h to prevent FA desaturation and elongation. 10 μM palmostatin B (Palm B) was used to preserve S-acylated residues in cell lysis buffer. The proportion of PLD2 in raft fractions was calculated as the ratio of the sum of PLD2 in raft fractions (fractions 3–5) to the sum of PLD2 in all 11 fractions. Data are presented as mean ± SEM (n = 3 independent experiments). Statistical significance was assessed using unpaired two-tailed t-tests (B–C) or one-way ANOVA followed by Dunnett’s multiple comparisons test (D–E), with significance thresholds defined as follows: ns, not significant, P ≥ 0.05; ∗, P < 0.05; ∗∗, P < 0.01; ∗∗∗, P < 0.001.

    Article Snippet: Additional reagents and kits were obtained as follows: Cdc42 Activation Assay Kit (#8819, Cell Signaling Technology), Bradford protein assay (Bio-Rad, #5000006), Phalloidin–TRITC (Sigma, #P1951), tris(2-carboxyethyl)phosphine hydrochloride (TCEP; Sigma, #C4706), methoxypolyethylene glycol maleimide (mPEG–Mal; Sigma, #63187), N-ethylmaleimide (NEM; Sigma, #E3876), protease and phosphatase inhibitors (Sigma, #P8340; #P0044), ELOVL6 inhibitor ELOVL6-IN-4 (MedChemExpress, #HY-152947), and SCD1 inhibitor A939572 (Biofine, #37062).

    Techniques: Confocal Microscopy, Western Blot, Isolation, Expressing, Lysis, Two Tailed Test

    The impact of PLD2 S-acylation site mutation on its targeting to lipid rafts. A: Western blot analysis of lipid raft fractions isolated by sucrose density gradient ultracentrifugation from HEK293T cells expressing EGFP-PLD2 (WT, C223A, C224A, and C223AC224A). Cells were starved with serum-free medium and preincubated with 1 μM SCD1 inhibitor A939572 and 1 μM ELOVL6 inhibitor ELOVL6-IN-4 for 3 h to prevent FA desaturation and elongation. Cells were then treated with BSA (Ctrl groups) or FAs (100 μM OA for WT, C223A, C224A; 100 μM palmitate for C223AC224A) for 3 h. Fractions 3–5 pinpoint lipid raft-enriched fractions, as indicated by the presence of Caveolin-1. B: Quantification of the proportion of PLD2 in raft fractions relative to the total PLD2 across all fractions. C: Confocal microscopy images of HEK293T cells expressing EGFP-PLD2-WT and -C223AC224A. Cells were then stained with anti-PIP 2 antibody (conjugated with Alexa Fluor 647 secondary antibody) to assess colocalization with EGFP-PLD2. The scale bar in the first micrograph applies to all images in the series of panel. The zoomed-in channels correspond to the regions highlighted by the yellow rectangles in the merged image. Colocalization index was assessed using Pearson's correlation coefficient. Data are presented as mean ± SEM (n = 3 independent experiments). Statistical significances were analyzed using one-way ANOVA followed by Dunnett’s multiple comparisons test to WT in Ctrl group for (B). Unpaired two-tailed Student's t-tests were used in FA treated group to compare the FA treated group and the corresponding BSA-treated control for (B) and WT versus C223AC224A for (C). Significance thresholds were defined as: ns, not significant, P ≥ 0.05; ∗∗, P < 0.01; ∗∗∗∗, P < 0.0001.

    Journal: Journal of Lipid Research

    Article Title: Oleate activates PLD2 lipase and GEF activity by modulating membrane microdomain dynamics via S-acylation

    doi: 10.1016/j.jlr.2025.100939

    Figure Lengend Snippet: The impact of PLD2 S-acylation site mutation on its targeting to lipid rafts. A: Western blot analysis of lipid raft fractions isolated by sucrose density gradient ultracentrifugation from HEK293T cells expressing EGFP-PLD2 (WT, C223A, C224A, and C223AC224A). Cells were starved with serum-free medium and preincubated with 1 μM SCD1 inhibitor A939572 and 1 μM ELOVL6 inhibitor ELOVL6-IN-4 for 3 h to prevent FA desaturation and elongation. Cells were then treated with BSA (Ctrl groups) or FAs (100 μM OA for WT, C223A, C224A; 100 μM palmitate for C223AC224A) for 3 h. Fractions 3–5 pinpoint lipid raft-enriched fractions, as indicated by the presence of Caveolin-1. B: Quantification of the proportion of PLD2 in raft fractions relative to the total PLD2 across all fractions. C: Confocal microscopy images of HEK293T cells expressing EGFP-PLD2-WT and -C223AC224A. Cells were then stained with anti-PIP 2 antibody (conjugated with Alexa Fluor 647 secondary antibody) to assess colocalization with EGFP-PLD2. The scale bar in the first micrograph applies to all images in the series of panel. The zoomed-in channels correspond to the regions highlighted by the yellow rectangles in the merged image. Colocalization index was assessed using Pearson's correlation coefficient. Data are presented as mean ± SEM (n = 3 independent experiments). Statistical significances were analyzed using one-way ANOVA followed by Dunnett’s multiple comparisons test to WT in Ctrl group for (B). Unpaired two-tailed Student's t-tests were used in FA treated group to compare the FA treated group and the corresponding BSA-treated control for (B) and WT versus C223AC224A for (C). Significance thresholds were defined as: ns, not significant, P ≥ 0.05; ∗∗, P < 0.01; ∗∗∗∗, P < 0.0001.

    Article Snippet: Additional reagents and kits were obtained as follows: Cdc42 Activation Assay Kit (#8819, Cell Signaling Technology), Bradford protein assay (Bio-Rad, #5000006), Phalloidin–TRITC (Sigma, #P1951), tris(2-carboxyethyl)phosphine hydrochloride (TCEP; Sigma, #C4706), methoxypolyethylene glycol maleimide (mPEG–Mal; Sigma, #63187), N-ethylmaleimide (NEM; Sigma, #E3876), protease and phosphatase inhibitors (Sigma, #P8340; #P0044), ELOVL6 inhibitor ELOVL6-IN-4 (MedChemExpress, #HY-152947), and SCD1 inhibitor A939572 (Biofine, #37062).

    Techniques: Mutagenesis, Western Blot, Isolation, Expressing, Confocal Microscopy, Staining, Two Tailed Test, Control