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topfluor cholesterol  (Croda International Plc)

 
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    Structured Review

    Croda International Plc topfluor cholesterol
    (A) Representative confocal images of HeLa WT cells preloaded for 24h with either OA or a mix of OA and <t>cholesterol</t> in the indicated concentrations, followed by a 1 h pulse with OA containing 1 mol% BODIPY-OA (Bpy-OA). Polarized light and BODIPY staining reveal distinct LD subpopulations with isotropic or LC internal organization. Nuclei were stained with Hoechst. Experiments were repeated three times with similar results. Scale bars, 5 µm; insets, 1 µm. (B) Quantification of Bpy-OA enrichment in LDs under the conditions described in (A). Enrichment of Bpy-TG was calculated as the ratio between LD and ER membrane fluorescence signals. Data are presented as SuperPlots, where small dots represent the mean value of one cell; large dots indicate the mean of each independent biological replicate ( N = 3). LDs were separated into two groups for the equimolar condition (100 µM Chol + 100 µM OA): non-LC LDs and LC LDs. Schematic cartoons above the graphs illustrate the predominant LD organization (Isotropic vs LC). Bars show mean ± SD. Statistical significance was determined using Student’s t -test: 200 µM OA, n = 23 cells; 190 µM Chol + 10 µM OA, n = 35 cells, * p = 0.0273; 100 µM Chol + 100 µM OA non-LC LDs, n = 30 cells; LC LDs, n = 23 cells, * p = 0.0491. (C) Representative confocal images of HeLa WT cells treated as in (A) but pulsed for 1 h with cholesterol containing 1 <t>mol%</t> <t>TopFluor-cholesterol</t> (TopFluor-Chol) to monitor CE incorporation into LDs. Polarized light and LipidTox staining reveal distinct LD subpopulations with isotropic or LC internal organization. Nuclei were stained with Hoechst. Scale bars, 5 µm; insets, 1 µm. (D) Quantification of TopFluor-Chol enrichment in LDs under the three loading conditions. Enrichment was calculated as the ratio between LD and ER membrane signals. Data are presented as SuperPlots, where small dots represent the mean value of one cell; large dots indicate the mean of each independent biological replicate ( N = 3). LDs were separated into two groups for the equimolar condition: non-LC LDs and LC LDs. Schematic cartoons above the graphs illustrate the predominant LD organization (Isotropic vs LC). Bars show mean ± SD. Statistical significance was determined using Student’s t -test: 200 µM OA, n = 31 cells; 190 µM Chol + 10 µM OA, n = 20 cells; 100 µM Chol + 100 µM OA non-LC LDs, n = 53 cells; LC LDs, n = 35 cells, * p = 0.0438.
    Topfluor Cholesterol, supplied by Croda International Plc, used in various techniques. Bioz Stars score: 96/100, based on 314 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Images

    1) Product Images from "Triglyceride/Cholesterol Ester Ratio Encodes Lipid Droplet Size and Diversity"

    Article Title: Triglyceride/Cholesterol Ester Ratio Encodes Lipid Droplet Size and Diversity

    Journal: bioRxiv

    doi: 10.64898/2026.01.21.700800

    (A) Representative confocal images of HeLa WT cells preloaded for 24h with either OA or a mix of OA and cholesterol in the indicated concentrations, followed by a 1 h pulse with OA containing 1 mol% BODIPY-OA (Bpy-OA). Polarized light and BODIPY staining reveal distinct LD subpopulations with isotropic or LC internal organization. Nuclei were stained with Hoechst. Experiments were repeated three times with similar results. Scale bars, 5 µm; insets, 1 µm. (B) Quantification of Bpy-OA enrichment in LDs under the conditions described in (A). Enrichment of Bpy-TG was calculated as the ratio between LD and ER membrane fluorescence signals. Data are presented as SuperPlots, where small dots represent the mean value of one cell; large dots indicate the mean of each independent biological replicate ( N = 3). LDs were separated into two groups for the equimolar condition (100 µM Chol + 100 µM OA): non-LC LDs and LC LDs. Schematic cartoons above the graphs illustrate the predominant LD organization (Isotropic vs LC). Bars show mean ± SD. Statistical significance was determined using Student’s t -test: 200 µM OA, n = 23 cells; 190 µM Chol + 10 µM OA, n = 35 cells, * p = 0.0273; 100 µM Chol + 100 µM OA non-LC LDs, n = 30 cells; LC LDs, n = 23 cells, * p = 0.0491. (C) Representative confocal images of HeLa WT cells treated as in (A) but pulsed for 1 h with cholesterol containing 1 mol% TopFluor-cholesterol (TopFluor-Chol) to monitor CE incorporation into LDs. Polarized light and LipidTox staining reveal distinct LD subpopulations with isotropic or LC internal organization. Nuclei were stained with Hoechst. Scale bars, 5 µm; insets, 1 µm. (D) Quantification of TopFluor-Chol enrichment in LDs under the three loading conditions. Enrichment was calculated as the ratio between LD and ER membrane signals. Data are presented as SuperPlots, where small dots represent the mean value of one cell; large dots indicate the mean of each independent biological replicate ( N = 3). LDs were separated into two groups for the equimolar condition: non-LC LDs and LC LDs. Schematic cartoons above the graphs illustrate the predominant LD organization (Isotropic vs LC). Bars show mean ± SD. Statistical significance was determined using Student’s t -test: 200 µM OA, n = 31 cells; 190 µM Chol + 10 µM OA, n = 20 cells; 100 µM Chol + 100 µM OA non-LC LDs, n = 53 cells; LC LDs, n = 35 cells, * p = 0.0438.
    Figure Legend Snippet: (A) Representative confocal images of HeLa WT cells preloaded for 24h with either OA or a mix of OA and cholesterol in the indicated concentrations, followed by a 1 h pulse with OA containing 1 mol% BODIPY-OA (Bpy-OA). Polarized light and BODIPY staining reveal distinct LD subpopulations with isotropic or LC internal organization. Nuclei were stained with Hoechst. Experiments were repeated three times with similar results. Scale bars, 5 µm; insets, 1 µm. (B) Quantification of Bpy-OA enrichment in LDs under the conditions described in (A). Enrichment of Bpy-TG was calculated as the ratio between LD and ER membrane fluorescence signals. Data are presented as SuperPlots, where small dots represent the mean value of one cell; large dots indicate the mean of each independent biological replicate ( N = 3). LDs were separated into two groups for the equimolar condition (100 µM Chol + 100 µM OA): non-LC LDs and LC LDs. Schematic cartoons above the graphs illustrate the predominant LD organization (Isotropic vs LC). Bars show mean ± SD. Statistical significance was determined using Student’s t -test: 200 µM OA, n = 23 cells; 190 µM Chol + 10 µM OA, n = 35 cells, * p = 0.0273; 100 µM Chol + 100 µM OA non-LC LDs, n = 30 cells; LC LDs, n = 23 cells, * p = 0.0491. (C) Representative confocal images of HeLa WT cells treated as in (A) but pulsed for 1 h with cholesterol containing 1 mol% TopFluor-cholesterol (TopFluor-Chol) to monitor CE incorporation into LDs. Polarized light and LipidTox staining reveal distinct LD subpopulations with isotropic or LC internal organization. Nuclei were stained with Hoechst. Scale bars, 5 µm; insets, 1 µm. (D) Quantification of TopFluor-Chol enrichment in LDs under the three loading conditions. Enrichment was calculated as the ratio between LD and ER membrane signals. Data are presented as SuperPlots, where small dots represent the mean value of one cell; large dots indicate the mean of each independent biological replicate ( N = 3). LDs were separated into two groups for the equimolar condition: non-LC LDs and LC LDs. Schematic cartoons above the graphs illustrate the predominant LD organization (Isotropic vs LC). Bars show mean ± SD. Statistical significance was determined using Student’s t -test: 200 µM OA, n = 31 cells; 190 µM Chol + 10 µM OA, n = 20 cells; 100 µM Chol + 100 µM OA non-LC LDs, n = 53 cells; LC LDs, n = 35 cells, * p = 0.0438.

    Techniques Used: Staining, Membrane, Fluorescence

    ( A-B ) HeLa WT cells were imaged after 24 hours of treatment with 200 µM Bpy-OA (A) or 50 µM TopFluor-Cholesterol (B), showing incorporation into the LDs and perfect colocalization with mature LD markers (Bodipy and LipidTox). Each experiment was repeated three times with similar results. (C) HeLa WT cells were imaged after 24 hours of loading with OA or a mixture of OA and cholesterol at the specified concentrations, followed by a 1-hour incubation with Bpy-OA. Experiments were repeated three times with consistent results. (D) HeLa WT cells were imaged after 24 hours of loading with OA or a mixture of OA and cholesterol at the indicated concentrations, followed by a 1-hour incubation with TopFluor-Cholesterol. Experiments were repeated three times with similar results.
    Figure Legend Snippet: ( A-B ) HeLa WT cells were imaged after 24 hours of treatment with 200 µM Bpy-OA (A) or 50 µM TopFluor-Cholesterol (B), showing incorporation into the LDs and perfect colocalization with mature LD markers (Bodipy and LipidTox). Each experiment was repeated three times with similar results. (C) HeLa WT cells were imaged after 24 hours of loading with OA or a mixture of OA and cholesterol at the specified concentrations, followed by a 1-hour incubation with Bpy-OA. Experiments were repeated three times with consistent results. (D) HeLa WT cells were imaged after 24 hours of loading with OA or a mixture of OA and cholesterol at the indicated concentrations, followed by a 1-hour incubation with TopFluor-Cholesterol. Experiments were repeated three times with similar results.

    Techniques Used: Incubation

    (A) Schematic illustration of labeled neutral lipid synthesis without seipin. SKO cells were loaded with BODIPY-OA or TopFluor-Cholesterol for 1 hour, then synthesized BODIPY-triacylglycerol (Bpy-TG) or TopFluor-Cholesteryl ester (TF-CE), respectively. The incorporation of Bpy-TG or TF-CE into existing non-LC and LC lipid droplets (LDs) was measured in the absence of seipin. (B) HeLa SKO cells were imaged after 24 hours of loading with OA, or a mixture of OA and cholesterol at specified concentrations, followed by a 1-hour incubation with Bpy-OA or TF-Chol, as indicated. Experiments were repeated three times with similar results. (C) Analysis of the conditions with 200 µM OA and 190 µM Chol + 10 µM OA from (B). Enrichment of Bpy-TG in the LDs was calculated as a ratio of the LDs’ signals to the membrane’s signals. Data are displayed as SuperPlots, with each point representing the mean from a single cell; large dots denote the mean of three independent replicates ( N = 3). Statistical significance was tested using Student’s t -test for the 200 µM OA condition, and Mann–Whitney test for the 190 µM Chol + 10 µM OA condition: WT n = 23 cells, SKO n = 32 cells for 200 µM OA, with p = 0.0825; WT n = 35 cells, SKO n = 33 cells for 190 µM Chol + 10 µM OA, with p = 0.1666. (D) Analysis of the same conditions from (B), focusing on TF-CE enrichment in the LDs, which was calculated as a ratio of LDs to membrane signals. Each small dot marks the mean for a single cell; large dots indicate the mean of three independent replicates ( N = 3). Statistical analysis used Student’s t -test or Mann–Whitney test: WT n = 31 cells, SKO n = 33 cells for 200 µM OA; WT n = 20, SKO n = 19 for 190 µM Chol + 10 µM OA, with p = 0.9081. (E) Analysis of the 100 µM Chol + 100 µM OA condition from (B). Enrichment of Bpy-TG was determined as a ratio of LD to membrane signals. LDs were categorized into non-LC and LC groups. Bars represent mean ± SD. Mann–Whitney test was used for significance: non-LC LDs, WT n = 30, SKO n = 45; LC LDs, WT n = 23, SKO n = 28; with p = 0.0064. (F) Analysis of the 100 µM OA + 100 µM Chol condition from (B), measuring TF-CE enrichment as a ratio of LD to membrane signals. LDs were divided into non-LC and LC groups. Bars show mean ± SD. Mann– Whitney test results: WT n = 53, 35; SKO n = 43, 18 cells for non-LC a n d LC LDs, respectively.
    Figure Legend Snippet: (A) Schematic illustration of labeled neutral lipid synthesis without seipin. SKO cells were loaded with BODIPY-OA or TopFluor-Cholesterol for 1 hour, then synthesized BODIPY-triacylglycerol (Bpy-TG) or TopFluor-Cholesteryl ester (TF-CE), respectively. The incorporation of Bpy-TG or TF-CE into existing non-LC and LC lipid droplets (LDs) was measured in the absence of seipin. (B) HeLa SKO cells were imaged after 24 hours of loading with OA, or a mixture of OA and cholesterol at specified concentrations, followed by a 1-hour incubation with Bpy-OA or TF-Chol, as indicated. Experiments were repeated three times with similar results. (C) Analysis of the conditions with 200 µM OA and 190 µM Chol + 10 µM OA from (B). Enrichment of Bpy-TG in the LDs was calculated as a ratio of the LDs’ signals to the membrane’s signals. Data are displayed as SuperPlots, with each point representing the mean from a single cell; large dots denote the mean of three independent replicates ( N = 3). Statistical significance was tested using Student’s t -test for the 200 µM OA condition, and Mann–Whitney test for the 190 µM Chol + 10 µM OA condition: WT n = 23 cells, SKO n = 32 cells for 200 µM OA, with p = 0.0825; WT n = 35 cells, SKO n = 33 cells for 190 µM Chol + 10 µM OA, with p = 0.1666. (D) Analysis of the same conditions from (B), focusing on TF-CE enrichment in the LDs, which was calculated as a ratio of LDs to membrane signals. Each small dot marks the mean for a single cell; large dots indicate the mean of three independent replicates ( N = 3). Statistical analysis used Student’s t -test or Mann–Whitney test: WT n = 31 cells, SKO n = 33 cells for 200 µM OA; WT n = 20, SKO n = 19 for 190 µM Chol + 10 µM OA, with p = 0.9081. (E) Analysis of the 100 µM Chol + 100 µM OA condition from (B). Enrichment of Bpy-TG was determined as a ratio of LD to membrane signals. LDs were categorized into non-LC and LC groups. Bars represent mean ± SD. Mann–Whitney test was used for significance: non-LC LDs, WT n = 30, SKO n = 45; LC LDs, WT n = 23, SKO n = 28; with p = 0.0064. (F) Analysis of the 100 µM OA + 100 µM Chol condition from (B), measuring TF-CE enrichment as a ratio of LD to membrane signals. LDs were divided into non-LC and LC groups. Bars show mean ± SD. Mann– Whitney test results: WT n = 53, 35; SKO n = 43, 18 cells for non-LC a n d LC LDs, respectively.

    Techniques Used: Labeling, Synthesized, Incubation, MANN-WHITNEY, Membrane



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    (A) Representative confocal images of HeLa WT cells preloaded for 24h with either OA or a mix of OA and <t>cholesterol</t> in the indicated concentrations, followed by a 1 h pulse with OA containing 1 mol% BODIPY-OA (Bpy-OA). Polarized light and BODIPY staining reveal distinct LD subpopulations with isotropic or LC internal organization. Nuclei were stained with Hoechst. Experiments were repeated three times with similar results. Scale bars, 5 µm; insets, 1 µm. (B) Quantification of Bpy-OA enrichment in LDs under the conditions described in (A). Enrichment of Bpy-TG was calculated as the ratio between LD and ER membrane fluorescence signals. Data are presented as SuperPlots, where small dots represent the mean value of one cell; large dots indicate the mean of each independent biological replicate ( N = 3). LDs were separated into two groups for the equimolar condition (100 µM Chol + 100 µM OA): non-LC LDs and LC LDs. Schematic cartoons above the graphs illustrate the predominant LD organization (Isotropic vs LC). Bars show mean ± SD. Statistical significance was determined using Student’s t -test: 200 µM OA, n = 23 cells; 190 µM Chol + 10 µM OA, n = 35 cells, * p = 0.0273; 100 µM Chol + 100 µM OA non-LC LDs, n = 30 cells; LC LDs, n = 23 cells, * p = 0.0491. (C) Representative confocal images of HeLa WT cells treated as in (A) but pulsed for 1 h with cholesterol containing 1 <t>mol%</t> <t>TopFluor-cholesterol</t> (TopFluor-Chol) to monitor CE incorporation into LDs. Polarized light and LipidTox staining reveal distinct LD subpopulations with isotropic or LC internal organization. Nuclei were stained with Hoechst. Scale bars, 5 µm; insets, 1 µm. (D) Quantification of TopFluor-Chol enrichment in LDs under the three loading conditions. Enrichment was calculated as the ratio between LD and ER membrane signals. Data are presented as SuperPlots, where small dots represent the mean value of one cell; large dots indicate the mean of each independent biological replicate ( N = 3). LDs were separated into two groups for the equimolar condition: non-LC LDs and LC LDs. Schematic cartoons above the graphs illustrate the predominant LD organization (Isotropic vs LC). Bars show mean ± SD. Statistical significance was determined using Student’s t -test: 200 µM OA, n = 31 cells; 190 µM Chol + 10 µM OA, n = 20 cells; 100 µM Chol + 100 µM OA non-LC LDs, n = 53 cells; LC LDs, n = 35 cells, * p = 0.0438.
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    Croda International Plc norchol
    Targeting and Internalization studies. A) Expression of FR-αβ analyzed by flow cytometry in the indicated cell lines. / B) Expression of FR-αβ analyzed by flow cytometry in isolated cells from xenografts in nude mice. / Percentage of Capan-2 cells that have internalized the pDNA LNPs ( C ) or mRNA LNPs ( D ), analyzed by flow cytometry. / E) Confocal microscopy images of internalization and endosomal escape of LNP-pDNA-FA-aCD47 (i) and LNP-mRNA-FA-aCD47 (ii) in Capan-2 cells after 24 h. Nuclei marker (Hoechst) in blue; lysosome marker LysoTracker™ Deep Red and membrane marker WGA Alexa Fluor 633 in red; and LNPs marked with <t>TopFluor®</t> <t>cholesterol,</t> with fluorescein labelled pDNA or with Andy Fluor 488 labelled mRNA are displayed in green. Scale bar: 20 µm; enlargements 10 µm. In each composition bottom right includes 2D scatterplot of colocalization and Person’s colocalization coefficient (Rr*) between the green signal (genetic material) and the red signal (lysosomes) of the bottom left image. Data are expressed as the means ± SEM of n = 3–7 independent replicates. Statistical analysis with Mann-Whitney test in panels A-B and a one-way ordinary ANOVA (Tukey’s multiple comparisons post-hoc) in panels C-D. *p < 0.05; **p < 0.01; ***p < 0.001. FR-αβ: folate receptor; RLU: relative light units; LNP: lipid nanoparticle; FA: folate; aCD47: anti-CD47 antibody .
    Norchol, supplied by Croda International Plc, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/norchol/product/Croda International Plc
    Average 96 stars, based on 1 article reviews
    norchol - by Bioz Stars, 2026-02
    96/100 stars
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    dipyrrometheneboron difluoride 24 norcholesterol topfluor chol  (Croda International Plc)
    96
    Croda International Plc dipyrrometheneboron difluoride 24 norcholesterol topfluor chol
    Targeting and Internalization studies. A) Expression of FR-αβ analyzed by flow cytometry in the indicated cell lines. / B) Expression of FR-αβ analyzed by flow cytometry in isolated cells from xenografts in nude mice. / Percentage of Capan-2 cells that have internalized the pDNA LNPs ( C ) or mRNA LNPs ( D ), analyzed by flow cytometry. / E) Confocal microscopy images of internalization and endosomal escape of LNP-pDNA-FA-aCD47 (i) and LNP-mRNA-FA-aCD47 (ii) in Capan-2 cells after 24 h. Nuclei marker (Hoechst) in blue; lysosome marker LysoTracker™ Deep Red and membrane marker WGA Alexa Fluor 633 in red; and LNPs marked with <t>TopFluor®</t> <t>cholesterol,</t> with fluorescein labelled pDNA or with Andy Fluor 488 labelled mRNA are displayed in green. Scale bar: 20 µm; enlargements 10 µm. In each composition bottom right includes 2D scatterplot of colocalization and Person’s colocalization coefficient (Rr*) between the green signal (genetic material) and the red signal (lysosomes) of the bottom left image. Data are expressed as the means ± SEM of n = 3–7 independent replicates. Statistical analysis with Mann-Whitney test in panels A-B and a one-way ordinary ANOVA (Tukey’s multiple comparisons post-hoc) in panels C-D. *p < 0.05; **p < 0.01; ***p < 0.001. FR-αβ: folate receptor; RLU: relative light units; LNP: lipid nanoparticle; FA: folate; aCD47: anti-CD47 antibody .
    Dipyrrometheneboron Difluoride 24 Norcholesterol Topfluor Chol, supplied by Croda International Plc, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/dipyrrometheneboron difluoride 24 norcholesterol topfluor chol/product/Croda International Plc
    Average 96 stars, based on 1 article reviews
    dipyrrometheneboron difluoride 24 norcholesterol topfluor chol - by Bioz Stars, 2026-02
    96/100 stars
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    lo marker topfluor cholesterol  (Croda International Plc)
    96
    Croda International Plc lo marker topfluor cholesterol
    Targeting and Internalization studies. A) Expression of FR-αβ analyzed by flow cytometry in the indicated cell lines. / B) Expression of FR-αβ analyzed by flow cytometry in isolated cells from xenografts in nude mice. / Percentage of Capan-2 cells that have internalized the pDNA LNPs ( C ) or mRNA LNPs ( D ), analyzed by flow cytometry. / E) Confocal microscopy images of internalization and endosomal escape of LNP-pDNA-FA-aCD47 (i) and LNP-mRNA-FA-aCD47 (ii) in Capan-2 cells after 24 h. Nuclei marker (Hoechst) in blue; lysosome marker LysoTracker™ Deep Red and membrane marker WGA Alexa Fluor 633 in red; and LNPs marked with <t>TopFluor®</t> <t>cholesterol,</t> with fluorescein labelled pDNA or with Andy Fluor 488 labelled mRNA are displayed in green. Scale bar: 20 µm; enlargements 10 µm. In each composition bottom right includes 2D scatterplot of colocalization and Person’s colocalization coefficient (Rr*) between the green signal (genetic material) and the red signal (lysosomes) of the bottom left image. Data are expressed as the means ± SEM of n = 3–7 independent replicates. Statistical analysis with Mann-Whitney test in panels A-B and a one-way ordinary ANOVA (Tukey’s multiple comparisons post-hoc) in panels C-D. *p < 0.05; **p < 0.01; ***p < 0.001. FR-αβ: folate receptor; RLU: relative light units; LNP: lipid nanoparticle; FA: folate; aCD47: anti-CD47 antibody .
    Lo Marker Topfluor Cholesterol, supplied by Croda International Plc, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/lo marker topfluor cholesterol/product/Croda International Plc
    Average 96 stars, based on 1 article reviews
    lo marker topfluor cholesterol - by Bioz Stars, 2026-02
    96/100 stars
    		  Buy from Supplier

    Image Search Results


    (A) Representative confocal images of HeLa WT cells preloaded for 24h with either OA or a mix of OA and cholesterol in the indicated concentrations, followed by a 1 h pulse with OA containing 1 mol% BODIPY-OA (Bpy-OA). Polarized light and BODIPY staining reveal distinct LD subpopulations with isotropic or LC internal organization. Nuclei were stained with Hoechst. Experiments were repeated three times with similar results. Scale bars, 5 µm; insets, 1 µm. (B) Quantification of Bpy-OA enrichment in LDs under the conditions described in (A). Enrichment of Bpy-TG was calculated as the ratio between LD and ER membrane fluorescence signals. Data are presented as SuperPlots, where small dots represent the mean value of one cell; large dots indicate the mean of each independent biological replicate ( N = 3). LDs were separated into two groups for the equimolar condition (100 µM Chol + 100 µM OA): non-LC LDs and LC LDs. Schematic cartoons above the graphs illustrate the predominant LD organization (Isotropic vs LC). Bars show mean ± SD. Statistical significance was determined using Student’s t -test: 200 µM OA, n = 23 cells; 190 µM Chol + 10 µM OA, n = 35 cells, * p = 0.0273; 100 µM Chol + 100 µM OA non-LC LDs, n = 30 cells; LC LDs, n = 23 cells, * p = 0.0491. (C) Representative confocal images of HeLa WT cells treated as in (A) but pulsed for 1 h with cholesterol containing 1 mol% TopFluor-cholesterol (TopFluor-Chol) to monitor CE incorporation into LDs. Polarized light and LipidTox staining reveal distinct LD subpopulations with isotropic or LC internal organization. Nuclei were stained with Hoechst. Scale bars, 5 µm; insets, 1 µm. (D) Quantification of TopFluor-Chol enrichment in LDs under the three loading conditions. Enrichment was calculated as the ratio between LD and ER membrane signals. Data are presented as SuperPlots, where small dots represent the mean value of one cell; large dots indicate the mean of each independent biological replicate ( N = 3). LDs were separated into two groups for the equimolar condition: non-LC LDs and LC LDs. Schematic cartoons above the graphs illustrate the predominant LD organization (Isotropic vs LC). Bars show mean ± SD. Statistical significance was determined using Student’s t -test: 200 µM OA, n = 31 cells; 190 µM Chol + 10 µM OA, n = 20 cells; 100 µM Chol + 100 µM OA non-LC LDs, n = 53 cells; LC LDs, n = 35 cells, * p = 0.0438.

    Journal: bioRxiv

    Article Title: Triglyceride/Cholesterol Ester Ratio Encodes Lipid Droplet Size and Diversity

    doi: 10.64898/2026.01.21.700800

    Figure Lengend Snippet: (A) Representative confocal images of HeLa WT cells preloaded for 24h with either OA or a mix of OA and cholesterol in the indicated concentrations, followed by a 1 h pulse with OA containing 1 mol% BODIPY-OA (Bpy-OA). Polarized light and BODIPY staining reveal distinct LD subpopulations with isotropic or LC internal organization. Nuclei were stained with Hoechst. Experiments were repeated three times with similar results. Scale bars, 5 µm; insets, 1 µm. (B) Quantification of Bpy-OA enrichment in LDs under the conditions described in (A). Enrichment of Bpy-TG was calculated as the ratio between LD and ER membrane fluorescence signals. Data are presented as SuperPlots, where small dots represent the mean value of one cell; large dots indicate the mean of each independent biological replicate ( N = 3). LDs were separated into two groups for the equimolar condition (100 µM Chol + 100 µM OA): non-LC LDs and LC LDs. Schematic cartoons above the graphs illustrate the predominant LD organization (Isotropic vs LC). Bars show mean ± SD. Statistical significance was determined using Student’s t -test: 200 µM OA, n = 23 cells; 190 µM Chol + 10 µM OA, n = 35 cells, * p = 0.0273; 100 µM Chol + 100 µM OA non-LC LDs, n = 30 cells; LC LDs, n = 23 cells, * p = 0.0491. (C) Representative confocal images of HeLa WT cells treated as in (A) but pulsed for 1 h with cholesterol containing 1 mol% TopFluor-cholesterol (TopFluor-Chol) to monitor CE incorporation into LDs. Polarized light and LipidTox staining reveal distinct LD subpopulations with isotropic or LC internal organization. Nuclei were stained with Hoechst. Scale bars, 5 µm; insets, 1 µm. (D) Quantification of TopFluor-Chol enrichment in LDs under the three loading conditions. Enrichment was calculated as the ratio between LD and ER membrane signals. Data are presented as SuperPlots, where small dots represent the mean value of one cell; large dots indicate the mean of each independent biological replicate ( N = 3). LDs were separated into two groups for the equimolar condition: non-LC LDs and LC LDs. Schematic cartoons above the graphs illustrate the predominant LD organization (Isotropic vs LC). Bars show mean ± SD. Statistical significance was determined using Student’s t -test: 200 µM OA, n = 31 cells; 190 µM Chol + 10 µM OA, n = 20 cells; 100 µM Chol + 100 µM OA non-LC LDs, n = 53 cells; LC LDs, n = 35 cells, * p = 0.0438.

    Article Snippet: For the preparation of labeled cholesterol (TopFluor-Chol), a mixture of TopFluor® Cholesterol (810255 P, Avanti Polar Lipids) with pure cholesterol (as described above) at a 1/100 molar ratio (TopFluor cholesterol/cholesterol) was made before complexation with methyl-ß-cyclodextrin.

    Techniques: Staining, Membrane, Fluorescence

    ( A-B ) HeLa WT cells were imaged after 24 hours of treatment with 200 µM Bpy-OA (A) or 50 µM TopFluor-Cholesterol (B), showing incorporation into the LDs and perfect colocalization with mature LD markers (Bodipy and LipidTox). Each experiment was repeated three times with similar results. (C) HeLa WT cells were imaged after 24 hours of loading with OA or a mixture of OA and cholesterol at the specified concentrations, followed by a 1-hour incubation with Bpy-OA. Experiments were repeated three times with consistent results. (D) HeLa WT cells were imaged after 24 hours of loading with OA or a mixture of OA and cholesterol at the indicated concentrations, followed by a 1-hour incubation with TopFluor-Cholesterol. Experiments were repeated three times with similar results.

    Journal: bioRxiv

    Article Title: Triglyceride/Cholesterol Ester Ratio Encodes Lipid Droplet Size and Diversity

    doi: 10.64898/2026.01.21.700800

    Figure Lengend Snippet: ( A-B ) HeLa WT cells were imaged after 24 hours of treatment with 200 µM Bpy-OA (A) or 50 µM TopFluor-Cholesterol (B), showing incorporation into the LDs and perfect colocalization with mature LD markers (Bodipy and LipidTox). Each experiment was repeated three times with similar results. (C) HeLa WT cells were imaged after 24 hours of loading with OA or a mixture of OA and cholesterol at the specified concentrations, followed by a 1-hour incubation with Bpy-OA. Experiments were repeated three times with consistent results. (D) HeLa WT cells were imaged after 24 hours of loading with OA or a mixture of OA and cholesterol at the indicated concentrations, followed by a 1-hour incubation with TopFluor-Cholesterol. Experiments were repeated three times with similar results.

    Article Snippet: For the preparation of labeled cholesterol (TopFluor-Chol), a mixture of TopFluor® Cholesterol (810255 P, Avanti Polar Lipids) with pure cholesterol (as described above) at a 1/100 molar ratio (TopFluor cholesterol/cholesterol) was made before complexation with methyl-ß-cyclodextrin.

    Techniques: Incubation

    (A) Schematic illustration of labeled neutral lipid synthesis without seipin. SKO cells were loaded with BODIPY-OA or TopFluor-Cholesterol for 1 hour, then synthesized BODIPY-triacylglycerol (Bpy-TG) or TopFluor-Cholesteryl ester (TF-CE), respectively. The incorporation of Bpy-TG or TF-CE into existing non-LC and LC lipid droplets (LDs) was measured in the absence of seipin. (B) HeLa SKO cells were imaged after 24 hours of loading with OA, or a mixture of OA and cholesterol at specified concentrations, followed by a 1-hour incubation with Bpy-OA or TF-Chol, as indicated. Experiments were repeated three times with similar results. (C) Analysis of the conditions with 200 µM OA and 190 µM Chol + 10 µM OA from (B). Enrichment of Bpy-TG in the LDs was calculated as a ratio of the LDs’ signals to the membrane’s signals. Data are displayed as SuperPlots, with each point representing the mean from a single cell; large dots denote the mean of three independent replicates ( N = 3). Statistical significance was tested using Student’s t -test for the 200 µM OA condition, and Mann–Whitney test for the 190 µM Chol + 10 µM OA condition: WT n = 23 cells, SKO n = 32 cells for 200 µM OA, with p = 0.0825; WT n = 35 cells, SKO n = 33 cells for 190 µM Chol + 10 µM OA, with p = 0.1666. (D) Analysis of the same conditions from (B), focusing on TF-CE enrichment in the LDs, which was calculated as a ratio of LDs to membrane signals. Each small dot marks the mean for a single cell; large dots indicate the mean of three independent replicates ( N = 3). Statistical analysis used Student’s t -test or Mann–Whitney test: WT n = 31 cells, SKO n = 33 cells for 200 µM OA; WT n = 20, SKO n = 19 for 190 µM Chol + 10 µM OA, with p = 0.9081. (E) Analysis of the 100 µM Chol + 100 µM OA condition from (B). Enrichment of Bpy-TG was determined as a ratio of LD to membrane signals. LDs were categorized into non-LC and LC groups. Bars represent mean ± SD. Mann–Whitney test was used for significance: non-LC LDs, WT n = 30, SKO n = 45; LC LDs, WT n = 23, SKO n = 28; with p = 0.0064. (F) Analysis of the 100 µM OA + 100 µM Chol condition from (B), measuring TF-CE enrichment as a ratio of LD to membrane signals. LDs were divided into non-LC and LC groups. Bars show mean ± SD. Mann– Whitney test results: WT n = 53, 35; SKO n = 43, 18 cells for non-LC a n d LC LDs, respectively.

    Journal: bioRxiv

    Article Title: Triglyceride/Cholesterol Ester Ratio Encodes Lipid Droplet Size and Diversity

    doi: 10.64898/2026.01.21.700800

    Figure Lengend Snippet: (A) Schematic illustration of labeled neutral lipid synthesis without seipin. SKO cells were loaded with BODIPY-OA or TopFluor-Cholesterol for 1 hour, then synthesized BODIPY-triacylglycerol (Bpy-TG) or TopFluor-Cholesteryl ester (TF-CE), respectively. The incorporation of Bpy-TG or TF-CE into existing non-LC and LC lipid droplets (LDs) was measured in the absence of seipin. (B) HeLa SKO cells were imaged after 24 hours of loading with OA, or a mixture of OA and cholesterol at specified concentrations, followed by a 1-hour incubation with Bpy-OA or TF-Chol, as indicated. Experiments were repeated three times with similar results. (C) Analysis of the conditions with 200 µM OA and 190 µM Chol + 10 µM OA from (B). Enrichment of Bpy-TG in the LDs was calculated as a ratio of the LDs’ signals to the membrane’s signals. Data are displayed as SuperPlots, with each point representing the mean from a single cell; large dots denote the mean of three independent replicates ( N = 3). Statistical significance was tested using Student’s t -test for the 200 µM OA condition, and Mann–Whitney test for the 190 µM Chol + 10 µM OA condition: WT n = 23 cells, SKO n = 32 cells for 200 µM OA, with p = 0.0825; WT n = 35 cells, SKO n = 33 cells for 190 µM Chol + 10 µM OA, with p = 0.1666. (D) Analysis of the same conditions from (B), focusing on TF-CE enrichment in the LDs, which was calculated as a ratio of LDs to membrane signals. Each small dot marks the mean for a single cell; large dots indicate the mean of three independent replicates ( N = 3). Statistical analysis used Student’s t -test or Mann–Whitney test: WT n = 31 cells, SKO n = 33 cells for 200 µM OA; WT n = 20, SKO n = 19 for 190 µM Chol + 10 µM OA, with p = 0.9081. (E) Analysis of the 100 µM Chol + 100 µM OA condition from (B). Enrichment of Bpy-TG was determined as a ratio of LD to membrane signals. LDs were categorized into non-LC and LC groups. Bars represent mean ± SD. Mann–Whitney test was used for significance: non-LC LDs, WT n = 30, SKO n = 45; LC LDs, WT n = 23, SKO n = 28; with p = 0.0064. (F) Analysis of the 100 µM OA + 100 µM Chol condition from (B), measuring TF-CE enrichment as a ratio of LD to membrane signals. LDs were divided into non-LC and LC groups. Bars show mean ± SD. Mann– Whitney test results: WT n = 53, 35; SKO n = 43, 18 cells for non-LC a n d LC LDs, respectively.

    Article Snippet: For the preparation of labeled cholesterol (TopFluor-Chol), a mixture of TopFluor® Cholesterol (810255 P, Avanti Polar Lipids) with pure cholesterol (as described above) at a 1/100 molar ratio (TopFluor cholesterol/cholesterol) was made before complexation with methyl-ß-cyclodextrin.

    Techniques: Labeling, Synthesized, Incubation, MANN-WHITNEY, Membrane

    Targeting and Internalization studies. A) Expression of FR-αβ analyzed by flow cytometry in the indicated cell lines. / B) Expression of FR-αβ analyzed by flow cytometry in isolated cells from xenografts in nude mice. / Percentage of Capan-2 cells that have internalized the pDNA LNPs ( C ) or mRNA LNPs ( D ), analyzed by flow cytometry. / E) Confocal microscopy images of internalization and endosomal escape of LNP-pDNA-FA-aCD47 (i) and LNP-mRNA-FA-aCD47 (ii) in Capan-2 cells after 24 h. Nuclei marker (Hoechst) in blue; lysosome marker LysoTracker™ Deep Red and membrane marker WGA Alexa Fluor 633 in red; and LNPs marked with TopFluor® cholesterol, with fluorescein labelled pDNA or with Andy Fluor 488 labelled mRNA are displayed in green. Scale bar: 20 µm; enlargements 10 µm. In each composition bottom right includes 2D scatterplot of colocalization and Person’s colocalization coefficient (Rr*) between the green signal (genetic material) and the red signal (lysosomes) of the bottom left image. Data are expressed as the means ± SEM of n = 3–7 independent replicates. Statistical analysis with Mann-Whitney test in panels A-B and a one-way ordinary ANOVA (Tukey’s multiple comparisons post-hoc) in panels C-D. *p < 0.05; **p < 0.01; ***p < 0.001. FR-αβ: folate receptor; RLU: relative light units; LNP: lipid nanoparticle; FA: folate; aCD47: anti-CD47 antibody .

    Journal: Biomedicine & Pharmacotherapy

    Article Title: Folate- and aCD47-functionalized lipid nanoparticles efficiently deliver mRNA and remodel pancreatic tumor microenvironment

    doi: 10.1016/j.biopha.2025.118919

    Figure Lengend Snippet: Targeting and Internalization studies. A) Expression of FR-αβ analyzed by flow cytometry in the indicated cell lines. / B) Expression of FR-αβ analyzed by flow cytometry in isolated cells from xenografts in nude mice. / Percentage of Capan-2 cells that have internalized the pDNA LNPs ( C ) or mRNA LNPs ( D ), analyzed by flow cytometry. / E) Confocal microscopy images of internalization and endosomal escape of LNP-pDNA-FA-aCD47 (i) and LNP-mRNA-FA-aCD47 (ii) in Capan-2 cells after 24 h. Nuclei marker (Hoechst) in blue; lysosome marker LysoTracker™ Deep Red and membrane marker WGA Alexa Fluor 633 in red; and LNPs marked with TopFluor® cholesterol, with fluorescein labelled pDNA or with Andy Fluor 488 labelled mRNA are displayed in green. Scale bar: 20 µm; enlargements 10 µm. In each composition bottom right includes 2D scatterplot of colocalization and Person’s colocalization coefficient (Rr*) between the green signal (genetic material) and the red signal (lysosomes) of the bottom left image. Data are expressed as the means ± SEM of n = 3–7 independent replicates. Statistical analysis with Mann-Whitney test in panels A-B and a one-way ordinary ANOVA (Tukey’s multiple comparisons post-hoc) in panels C-D. *p < 0.05; **p < 0.01; ***p < 0.001. FR-αβ: folate receptor; RLU: relative light units; LNP: lipid nanoparticle; FA: folate; aCD47: anti-CD47 antibody .

    Article Snippet: The lipids 1-stearoyl-2-oleoyl-sn-glycero-3-phosphocholine (SOPC); 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP); (R)-2,3-bis(octadecyloxy)propyl-1-(methoxy polyethylene glycol 2000) carbamate (DMG-PEG(2000)); 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[folate(polyethylene glycol)-2000] (DSPE-PEG(2000)-FA); 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[amino(polyethylene glycol)-2000] (DSPE-PEG(2000)-Amine); 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine-N-(Cyanine 7) (18:1 Cy7 PE); and 23-(dipyrrometheneboron difluoride)-24-norcholesterol (TopFluor® Cholesterol) were purchased from Avanti Polar Lipids (Alabaster, AL).

    Techniques: Expressing, Flow Cytometry, Isolation, Confocal Microscopy, Marker, Membrane, MANN-WHITNEY