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daclatasvir (TargetMol)

Name: daclatasvir
Brand: TargetMol
Rating: 94 (5 reviews)

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TargetMol daclatasvir
Daclatasvir, supplied by TargetMol, used in various techniques. Bioz Stars score: 94/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/daclatasvir/product/TargetMol
Average 94 stars, based on 5 article reviews

daclatasvir - by Bioz Stars, 2026-02

94/100 stars

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Volcano plots of phenotype scores and P values from genome-wide CRISPR interference screens under treatment with ( A ) pralatrexate, ( C ) bortezomib, ( D ) gemcitabine, ( E ) romidepsin, and ( F ) oxaliplatin. Orange points denote gene knockdowns with false discovery rate (FDR) below 10%, calculated from comparison with ‘pseudogene’ controls from non-targeting guide RNAs (gray; Methods); this threshold corresponds to Mann Whitney P values <0.05 (noting that FDR is not type 1 error). B . Effect of SLC19A1 knockdown on pralatrexate resistance. Pralatrexate dose response was measured in SU-DHL-1 cells expressing dCas9-KRAB, either untransformed, transformed with a non-targeting guide RNA, or transformed with a guide RNA targeting SLC19A1. Error bars show the standard deviation of four replicate measurements. At a physiologically relevant pralatrexate concentration (60nM, based on human pharmacokinetic studies) SLC19A1 knockdown abolished pralatrexate response compared with non-targeting control ( P = 0.00002, Student’s t-test).

Journal: bioRxiv

Article Title: Integrating functional genomics and proteomics identifies Folate Carrier SLC19A1 as a predictor of pralatrexate sensitivity in T-cell lymphoma

doi: 10.1101/2025.10.08.681217

Figure Lengend Snippet: Volcano plots of phenotype scores and P values from genome-wide CRISPR interference screens under treatment with ( A ) pralatrexate, ( C ) bortezomib, ( D ) gemcitabine, ( E ) romidepsin, and ( F ) oxaliplatin. Orange points denote gene knockdowns with false discovery rate (FDR) below 10%, calculated from comparison with ‘pseudogene’ controls from non-targeting guide RNAs (gray; Methods); this threshold corresponds to Mann Whitney P values <0.05 (noting that FDR is not type 1 error). B . Effect of SLC19A1 knockdown on pralatrexate resistance. Pralatrexate dose response was measured in SU-DHL-1 cells expressing dCas9-KRAB, either untransformed, transformed with a non-targeting guide RNA, or transformed with a guide RNA targeting SLC19A1. Error bars show the standard deviation of four replicate measurements. At a physiologically relevant pralatrexate concentration (60nM, based on human pharmacokinetic studies) SLC19A1 knockdown abolished pralatrexate response compared with non-targeting control ( P = 0.00002, Student’s t-test).

Article Snippet: Five therapies used in the treatment of T-cell Lymphoma were purchased from MedChemExpress; romidepsin (Catalog number: HY15149, pralatrexate (Catalog number: HY-10466), bortezomib (Catalog Number: HY-10227), gemcitabine (Catalog Number: HY-17026), oxaliplatin (Catalog number: HY-17371).

Techniques: Genome Wide, CRISPR, Comparison, MANN-WHITNEY, Knockdown, Expressing, Transformation Assay, Standard Deviation, Concentration Assay, Control

A. Example dose-response measurements of T– and NK-cell lymphoma cultures treated with pralatrexate for 72 hours. Cell viability was measured by luminescence (CellTiter-Glo) relative to untreated controls. Replicates included two independently propagated cultures, assayed on different plates across different weeks, yielding 3-4 replicates for each of 30 lymphoma cultures. B. Normalized drug sensitivities of 30 lymphoma cultures to 5 therapies used in the second-line treatment of PTCL. Sensitivities were quantified as the area over the dose-response curve (AOC), up to a clinically relevant maximum concentration (0.9 μM romidepsin , 0.06 μM pralatrexate , 0.8 μM bortezomib , 0.1 μM gemcitabine , and 4 μM oxaliplatin ). White asterisks mark cultures in the top quartile of sensitivity to each therapy. C. Heatmap indicating which lymphoma cultures are in the top quartile of sensitivity to one or more drugs.

Journal: bioRxiv

Article Title: Integrating functional genomics and proteomics identifies Folate Carrier SLC19A1 as a predictor of pralatrexate sensitivity in T-cell lymphoma

doi: 10.1101/2025.10.08.681217

Figure Lengend Snippet: A. Example dose-response measurements of T– and NK-cell lymphoma cultures treated with pralatrexate for 72 hours. Cell viability was measured by luminescence (CellTiter-Glo) relative to untreated controls. Replicates included two independently propagated cultures, assayed on different plates across different weeks, yielding 3-4 replicates for each of 30 lymphoma cultures. B. Normalized drug sensitivities of 30 lymphoma cultures to 5 therapies used in the second-line treatment of PTCL. Sensitivities were quantified as the area over the dose-response curve (AOC), up to a clinically relevant maximum concentration (0.9 μM romidepsin , 0.06 μM pralatrexate , 0.8 μM bortezomib , 0.1 μM gemcitabine , and 4 μM oxaliplatin ). White asterisks mark cultures in the top quartile of sensitivity to each therapy. C. Heatmap indicating which lymphoma cultures are in the top quartile of sensitivity to one or more drugs.

Techniques: Concentration Assay

Screening strategy and functional validation of candidate drugs for MASLD treatment. A: Schematic representation of the characterization and screening strategy employed for the FDA-approved drug library. B: Representative Nile red staining images and quantitative analysis of HuH-7 cell MASLD models in the stimulation of palmitic acid/oleic acid (PA/OA, 0.25 mM/0.5 mM) and in the treatment respectively with 13 candidate drugs for 18 h. Scale bar, 25 μm. Each group was compared with the PA/OA+dimethyl sulfoxide (DMSO) group. C: Heatmap of fold changes representing the mRNA levels of ATGL , MAGL , IL1β , and TNFα in HuH-7 cell MASLD models challenged with PA/OA (0.25 mM/0.5 mM) and treated with the candidate drugs for 18 h. The PA/OA+DMSO group was served as the control. D: Venn diagram showing the overlap of drugs that reduced lipid droplets in (B), those that increased the expression of ATGL and MAGL , and decreased IL1β and TNFα expression in (C). E: Chemical structure, CAS number, and simplified structure of daclatasvir. F: Cell viability following 24 h-treatment with daclatasvir at the indicated concentrations in HuH-7 cells. Each group was compared with the DMSO group (labeled as daclatasvir 0 μM on the figure). For (B), n = 4 independent biological replicates; for (C), n = 3 independent biological replicates; for (F), n = 6 independent biological replicates. Data are presented as mean ± SD; ∗ P < 0.05, ∗∗ P < 0.01; one-way ANOVA in (B, C, F). BSA, bovine serum albumin; ACV, acyclovir; DCV, daclatasvir; DSV, dasabuvir; FCV, famciclovir; GLE, glecaprevir; IDU, idarubicin; LVD, lamivudine; OBV, ombitasvir; PTV, paritaprevir; RBV, ribavirin; SFV, sofosbuvir; TAF, tenofovir alafenamide; TDF, tenofovir disoproxil fumarate.

Journal: Journal of Lipid Research

Article Title: Hepatoprotective drug screening identifies daclatasvir, a promising therapeutic candidate for MASLD by targeting PLIN2

doi: 10.1016/j.jlr.2025.100835

Figure Lengend Snippet: Screening strategy and functional validation of candidate drugs for MASLD treatment. A: Schematic representation of the characterization and screening strategy employed for the FDA-approved drug library. B: Representative Nile red staining images and quantitative analysis of HuH-7 cell MASLD models in the stimulation of palmitic acid/oleic acid (PA/OA, 0.25 mM/0.5 mM) and in the treatment respectively with 13 candidate drugs for 18 h. Scale bar, 25 μm. Each group was compared with the PA/OA+dimethyl sulfoxide (DMSO) group. C: Heatmap of fold changes representing the mRNA levels of ATGL , MAGL , IL1β , and TNFα in HuH-7 cell MASLD models challenged with PA/OA (0.25 mM/0.5 mM) and treated with the candidate drugs for 18 h. The PA/OA+DMSO group was served as the control. D: Venn diagram showing the overlap of drugs that reduced lipid droplets in (B), those that increased the expression of ATGL and MAGL , and decreased IL1β and TNFα expression in (C). E: Chemical structure, CAS number, and simplified structure of daclatasvir. F: Cell viability following 24 h-treatment with daclatasvir at the indicated concentrations in HuH-7 cells. Each group was compared with the DMSO group (labeled as daclatasvir 0 μM on the figure). For (B), n = 4 independent biological replicates; for (C), n = 3 independent biological replicates; for (F), n = 6 independent biological replicates. Data are presented as mean ± SD; ∗ P < 0.05, ∗∗ P < 0.01; one-way ANOVA in (B, C, F). BSA, bovine serum albumin; ACV, acyclovir; DCV, daclatasvir; DSV, dasabuvir; FCV, famciclovir; GLE, glecaprevir; IDU, idarubicin; LVD, lamivudine; OBV, ombitasvir; PTV, paritaprevir; RBV, ribavirin; SFV, sofosbuvir; TAF, tenofovir alafenamide; TDF, tenofovir disoproxil fumarate.

Article Snippet: After cell adhesion, 10 μM daclatasvir or an alternative FDA-approved drug from the library (#L1300, Selleckchem) was added to the culture medium, along with an equivalent volume of DMSO (D2650; Sigma) as a vehicle control.

Techniques: Functional Assay, Biomarker Discovery, Drug discovery, Staining, Control, Expressing, Labeling

Daclatasvir mitigates lipid accumulation and inflammation in MASLD cell models. A: Representative images of Nile Red staining and quantitative analysis of lipid accumulation in HuH-7 cells pretreated with daclatasvir for 6 h, followed by co-treatment with PA/OA (0.25 mM/0.5 mM) for 18 h. Scale bar, 25 μm. B: Representative images and quantification of lipid accumulation in HuH-7 cells first stimulated with PA/OA (0.25 mM/0.5 mM) for 6 h, followed by daclatasvir treatment for an additional 8–12 h. Scale bar, 25 μm. C: Triglyceride (TG) and total cholesterol (TC) levels in HuH-7 cells treated with or without daclatasvir after 18 h of PA/OA (0.25 mM/0.5 mM) stimulation. D and E: KEGG enrichment analysis showed pathways (D) and heatmap showed the expression of differentially regulated genes (E) associated with lipid metabolism and inflammation, in HuH-7 cells challenged with PA (0.25 mM) and treated with daclatasvir for 12 h. F: Quantitative polymerase chain reaction (qPCR) assay of ATGL , MAGL , ACOX1 , CPT1α , and PPARα in HuH-7 cells challenged with palmitic acid (PA, 0.25 mM) and treated with or without daclatasvir for 12 h. For (A, B), (D,E), n = 3 independent biological replicates; for (C, F), n = 6 independent biological replicates. Data are presented as mean ± SD; ∗ P < 0.05, ∗∗ P < 0.01; one-way ANOVA in (A), and Student’s t test in (C, F).

Journal: Journal of Lipid Research

Article Title: Hepatoprotective drug screening identifies daclatasvir, a promising therapeutic candidate for MASLD by targeting PLIN2

doi: 10.1016/j.jlr.2025.100835

Figure Lengend Snippet: Daclatasvir mitigates lipid accumulation and inflammation in MASLD cell models. A: Representative images of Nile Red staining and quantitative analysis of lipid accumulation in HuH-7 cells pretreated with daclatasvir for 6 h, followed by co-treatment with PA/OA (0.25 mM/0.5 mM) for 18 h. Scale bar, 25 μm. B: Representative images and quantification of lipid accumulation in HuH-7 cells first stimulated with PA/OA (0.25 mM/0.5 mM) for 6 h, followed by daclatasvir treatment for an additional 8–12 h. Scale bar, 25 μm. C: Triglyceride (TG) and total cholesterol (TC) levels in HuH-7 cells treated with or without daclatasvir after 18 h of PA/OA (0.25 mM/0.5 mM) stimulation. D and E: KEGG enrichment analysis showed pathways (D) and heatmap showed the expression of differentially regulated genes (E) associated with lipid metabolism and inflammation, in HuH-7 cells challenged with PA (0.25 mM) and treated with daclatasvir for 12 h. F: Quantitative polymerase chain reaction (qPCR) assay of ATGL , MAGL , ACOX1 , CPT1α , and PPARα in HuH-7 cells challenged with palmitic acid (PA, 0.25 mM) and treated with or without daclatasvir for 12 h. For (A, B), (D,E), n = 3 independent biological replicates; for (C, F), n = 6 independent biological replicates. Data are presented as mean ± SD; ∗ P < 0.05, ∗∗ P < 0.01; one-way ANOVA in (A), and Student’s t test in (C, F).

Techniques: Staining, Expressing, Real-time Polymerase Chain Reaction

Daclatasvir treatment mitigates metabolic and inflammatory dysregulation in HFHC-induced MASH. A: Schematic illustration of the experimental workflow for the daclatasvir treatment strategy in a normal chow (NC), or a high-fat, high-cholesterol (HFHC) diet-induced MASH mouse model. B: Body weight, liver weight, and the ratio of liver weight to body weight (LW/BW) were recorded for NC or HFHC diet-fed mice after 12 weeks of treatment with either control solution or daclatasvir. C: Representative images of H&E, Oil Red O, and picrosirius red (PSR) staining of liver samples from vehicle or daclatasvir treated mice at 16 weeks of NC or HFHC diet administration. Scale bar, 200 μm. D: Quantitative results for Oil Red O and PSR staining shown in (C). E and F: Serum TG and TC concentrations (E), as well as enzyme activities of serum hepatic transaminases (F) in vehicle- or daclatasvir-treated mice after 16 weeks of HFHC feeding. G and H: KEGG enrichment analysis showed pathways (G) and heatmap illustrated the differential gene expression (H) related to lipid metabolism, inflammation and fibrosis, in livers from HFHC diet-induced MASH model treated with vehicle or daclatasvir. I: The mRNA expression of Atgl , Magl , Acox1 , Cpt1α , and Pparα in liver tissues from HFHC diet-induced MASH mice following 12 weeks of daclatasvir treatment. For (B–F), and (I), n = 6 mice per group; for (G, H), n = 3 mice per group. Data are presented as mean ± SD; ∗ P < 0.05, ∗∗ P < 0.01; one-way ANOVA in (B), and Student’s t test in other statistics.

Journal: Journal of Lipid Research

Article Title: Hepatoprotective drug screening identifies daclatasvir, a promising therapeutic candidate for MASLD by targeting PLIN2

doi: 10.1016/j.jlr.2025.100835

Figure Lengend Snippet: Daclatasvir treatment mitigates metabolic and inflammatory dysregulation in HFHC-induced MASH. A: Schematic illustration of the experimental workflow for the daclatasvir treatment strategy in a normal chow (NC), or a high-fat, high-cholesterol (HFHC) diet-induced MASH mouse model. B: Body weight, liver weight, and the ratio of liver weight to body weight (LW/BW) were recorded for NC or HFHC diet-fed mice after 12 weeks of treatment with either control solution or daclatasvir. C: Representative images of H&E, Oil Red O, and picrosirius red (PSR) staining of liver samples from vehicle or daclatasvir treated mice at 16 weeks of NC or HFHC diet administration. Scale bar, 200 μm. D: Quantitative results for Oil Red O and PSR staining shown in (C). E and F: Serum TG and TC concentrations (E), as well as enzyme activities of serum hepatic transaminases (F) in vehicle- or daclatasvir-treated mice after 16 weeks of HFHC feeding. G and H: KEGG enrichment analysis showed pathways (G) and heatmap illustrated the differential gene expression (H) related to lipid metabolism, inflammation and fibrosis, in livers from HFHC diet-induced MASH model treated with vehicle or daclatasvir. I: The mRNA expression of Atgl , Magl , Acox1 , Cpt1α , and Pparα in liver tissues from HFHC diet-induced MASH mice following 12 weeks of daclatasvir treatment. For (B–F), and (I), n = 6 mice per group; for (G, H), n = 3 mice per group. Data are presented as mean ± SD; ∗ P < 0.05, ∗∗ P < 0.01; one-way ANOVA in (B), and Student’s t test in other statistics.

Techniques: Control, Staining, Gene Expression, Expressing

Daclatasvir treatment ameliorates liver damage and fibrosis in MCD diet-induced MASH mouse model. A: Schematic illustration of the experimental workflow for the daclatasvir treatment strategy in a methionine-choline deficient (MCD) diet-induced MASH mouse model. B: Liver weight, LW/BW, and LW/BMI were recorded for MCD diet-fed mice after 3 weeks of treatment with either vehicle or daclatasvir. C: Representative images of H&E, PSR, and CD11b staining of liver samples from vehicle or daclatasvir treated mice at 4 weeks of MCD diet administration. Scale bar, 200 μm for H&E and PSR; 100 μm for CD11b. D: Quantitative results for PSR and CD11b staining shown in (C). E: Enzyme activities of serum hepatic transaminases in MCD diet-fed mice after 3 weeks of treatment with either vehicle or daclatasvir. F–H: The mRNA expression of genes associated with lipid catabolism (F), inflammation (G), and fibrosis (H) in liver tissues from MCD diet-induced MASH mice following 3 weeks of daclatasvir or vehicle treatment. For (B–H), n = 6 mice per group. Data are presented as mean ± SD; ∗ P < 0.05, ∗∗ P < 0.01; Student’s t test for statistics.

Journal: Journal of Lipid Research

Article Title: Hepatoprotective drug screening identifies daclatasvir, a promising therapeutic candidate for MASLD by targeting PLIN2

doi: 10.1016/j.jlr.2025.100835

Figure Lengend Snippet: Daclatasvir treatment ameliorates liver damage and fibrosis in MCD diet-induced MASH mouse model. A: Schematic illustration of the experimental workflow for the daclatasvir treatment strategy in a methionine-choline deficient (MCD) diet-induced MASH mouse model. B: Liver weight, LW/BW, and LW/BMI were recorded for MCD diet-fed mice after 3 weeks of treatment with either vehicle or daclatasvir. C: Representative images of H&E, PSR, and CD11b staining of liver samples from vehicle or daclatasvir treated mice at 4 weeks of MCD diet administration. Scale bar, 200 μm for H&E and PSR; 100 μm for CD11b. D: Quantitative results for PSR and CD11b staining shown in (C). E: Enzyme activities of serum hepatic transaminases in MCD diet-fed mice after 3 weeks of treatment with either vehicle or daclatasvir. F–H: The mRNA expression of genes associated with lipid catabolism (F), inflammation (G), and fibrosis (H) in liver tissues from MCD diet-induced MASH mice following 3 weeks of daclatasvir or vehicle treatment. For (B–H), n = 6 mice per group. Data are presented as mean ± SD; ∗ P < 0.05, ∗∗ P < 0.01; Student’s t test for statistics.

Techniques: Staining, Expressing

Daclatasvir targets PLIN2 for degradation via ubiquitination. A: The experimental workflow for screening potential target protein of daclatasvir. B: Table shows molecular docking analysis of nine candidate proteins with daclatasvir. The XP GScore reflects the binding affinity, and the MM-GBSA dG Bind reflects the binding free energy alterations upon complex formation. C: Molecular docking model demonstrating the interaction between daclatasvir and PLIN2 protein. The upper right box is a zoomed-in display of the lower left box. Red and blue arrows point to daclatasvir and PLIN2 proteins, respectively. D: Isothermal titration calorimetry analysis that illustrating the thermodynamic profiles of connection between daclatasvir and PLIN2 protein. E and F: Western blot analysis of PLIN2 protein levels in primary mouse hepatocytes treated with DMSO or daclatasvir and challenged with BSA or PA (0.5 mM) for 12 h (E), and those in liver tissues from NC or HFHC diet-fed mice treated with vehicle or daclatasvir (F). Quantitative data are shown on the right, using β-ACTIN as the loading control. G and H: PLIN2 protein levels in HuH-7 cells treated with DMSO or chloroquine (Chlq, 25 μM) (G), and those treated with DMSO or MG132 (10 μM) (H), following 12 h of PA (0.5 mM) stimulation and 4 h of cycloheximide (CHX, 100 μM) exposure. Quantitative data are shown on the bottom, using β-ACTIN as the loading control. I: Ubiquitination levels of PLIN2 in HEK 293T cells transfected with HA-tagged PLIN2 and Myc-tagged ubiquitin plasmids, treated with DMSO or daclatasvir (10 μM) in the presence of MG132 (10 μM). J and K: Ubiquitination levels of K11-linked PLIN2 (J) and K48-linked PLIN2 (K) in HEK 293T cells transfected with HA-tagged PLIN2 and Myc-tagged K11-only ubiquitin (Myc-Ub-K11O) (J) or Myc-tagged K48-only ubiquitin (Myc-Ub-K48O) (K) plasmids, treated with DMSO or daclatasvir (10 μM) in the presence of MG132 (10 μM). For (E–G), n = 3 independent biological replicates. Data are presented as mean ± SD; ∗ P < 0.05, ∗∗ P < 0.01, n.s., not significant; one-way ANOVA for statistics.

Journal: Journal of Lipid Research

Article Title: Hepatoprotective drug screening identifies daclatasvir, a promising therapeutic candidate for MASLD by targeting PLIN2

doi: 10.1016/j.jlr.2025.100835

Figure Lengend Snippet: Daclatasvir targets PLIN2 for degradation via ubiquitination. A: The experimental workflow for screening potential target protein of daclatasvir. B: Table shows molecular docking analysis of nine candidate proteins with daclatasvir. The XP GScore reflects the binding affinity, and the MM-GBSA dG Bind reflects the binding free energy alterations upon complex formation. C: Molecular docking model demonstrating the interaction between daclatasvir and PLIN2 protein. The upper right box is a zoomed-in display of the lower left box. Red and blue arrows point to daclatasvir and PLIN2 proteins, respectively. D: Isothermal titration calorimetry analysis that illustrating the thermodynamic profiles of connection between daclatasvir and PLIN2 protein. E and F: Western blot analysis of PLIN2 protein levels in primary mouse hepatocytes treated with DMSO or daclatasvir and challenged with BSA or PA (0.5 mM) for 12 h (E), and those in liver tissues from NC or HFHC diet-fed mice treated with vehicle or daclatasvir (F). Quantitative data are shown on the right, using β-ACTIN as the loading control. G and H: PLIN2 protein levels in HuH-7 cells treated with DMSO or chloroquine (Chlq, 25 μM) (G), and those treated with DMSO or MG132 (10 μM) (H), following 12 h of PA (0.5 mM) stimulation and 4 h of cycloheximide (CHX, 100 μM) exposure. Quantitative data are shown on the bottom, using β-ACTIN as the loading control. I: Ubiquitination levels of PLIN2 in HEK 293T cells transfected with HA-tagged PLIN2 and Myc-tagged ubiquitin plasmids, treated with DMSO or daclatasvir (10 μM) in the presence of MG132 (10 μM). J and K: Ubiquitination levels of K11-linked PLIN2 (J) and K48-linked PLIN2 (K) in HEK 293T cells transfected with HA-tagged PLIN2 and Myc-tagged K11-only ubiquitin (Myc-Ub-K11O) (J) or Myc-tagged K48-only ubiquitin (Myc-Ub-K48O) (K) plasmids, treated with DMSO or daclatasvir (10 μM) in the presence of MG132 (10 μM). For (E–G), n = 3 independent biological replicates. Data are presented as mean ± SD; ∗ P < 0.05, ∗∗ P < 0.01, n.s., not significant; one-way ANOVA for statistics.

Techniques: Ubiquitin Proteomics, Binding Assay, Isothermal Titration Calorimetry, Western Blot, Control, Transfection

Daclatasvir enhances MARCH6-mediated PLIN2 ubiquitination to alleviate lipid accumulation and inflammation. A, B: coimmunoprecipitation (CoIP) assay of the interaction between MARCH6 and PLIN2 via immunoprecipitating HA-tagged PLIN2 (A) or Flag-tagged MARCH6 (B) in HEK 293T cells treated with DMSO or daclatasvir (10 μM) for 12 h following transfection with indicated plasmids. C and D: CoIP assay of the interaction between UBR1 and PLIN2 via immunoprecipitating HA-tagged PLIN2 (C) or Flag-tagged UBR1 (D) in HEK 293T cells treated with DMSO or daclatasvir (10 μM) for 12 h following transfection with the indicated plasmids. E: Ubiquitination levels of PLIN2 in HEK 293T cells transfected with HA-tagged PLIN2, Flag-tagged MARCH6, and Myc-tagged Ub-K11O, in the presence of MG132 (10 μM). F: Ubiquitination levels of PLIN2 in HEK 293T cells transfected with HA-tagged PLIN2, Flag-tagged MARCH6, and Myc-tagged Ub-K11O, treated with DMSO or daclatasvir (10 μM) in the presence of MG132 (10 μM). G: Representative Nile Red staining images of HuH-7 cells overexpressing MARCH6 or control plasmid and suffering 18 h of PA/OA (0.25 mM/0.5 mM) stimulation, along with quantitative analysis of lipid accumulation. Scale bar, 25 μm. H: The mRNA levels of TNFα , MCP1 , and IL1β at 12 h post-PA (0.5 mM) stimulation in HuH-7 cells overexpressing MARCH6 or control plasmid. For (H), n = 6 independent biological replicates; for the others, n = 3 independent biological replicates. Data are presented as mean ± SD; ∗ P < 0.05, ∗∗ P < 0.01; one-way ANOVA for statistics.

Journal: Journal of Lipid Research

Article Title: Hepatoprotective drug screening identifies daclatasvir, a promising therapeutic candidate for MASLD by targeting PLIN2

doi: 10.1016/j.jlr.2025.100835

Figure Lengend Snippet: Daclatasvir enhances MARCH6-mediated PLIN2 ubiquitination to alleviate lipid accumulation and inflammation. A, B: coimmunoprecipitation (CoIP) assay of the interaction between MARCH6 and PLIN2 via immunoprecipitating HA-tagged PLIN2 (A) or Flag-tagged MARCH6 (B) in HEK 293T cells treated with DMSO or daclatasvir (10 μM) for 12 h following transfection with indicated plasmids. C and D: CoIP assay of the interaction between UBR1 and PLIN2 via immunoprecipitating HA-tagged PLIN2 (C) or Flag-tagged UBR1 (D) in HEK 293T cells treated with DMSO or daclatasvir (10 μM) for 12 h following transfection with the indicated plasmids. E: Ubiquitination levels of PLIN2 in HEK 293T cells transfected with HA-tagged PLIN2, Flag-tagged MARCH6, and Myc-tagged Ub-K11O, in the presence of MG132 (10 μM). F: Ubiquitination levels of PLIN2 in HEK 293T cells transfected with HA-tagged PLIN2, Flag-tagged MARCH6, and Myc-tagged Ub-K11O, treated with DMSO or daclatasvir (10 μM) in the presence of MG132 (10 μM). G: Representative Nile Red staining images of HuH-7 cells overexpressing MARCH6 or control plasmid and suffering 18 h of PA/OA (0.25 mM/0.5 mM) stimulation, along with quantitative analysis of lipid accumulation. Scale bar, 25 μm. H: The mRNA levels of TNFα , MCP1 , and IL1β at 12 h post-PA (0.5 mM) stimulation in HuH-7 cells overexpressing MARCH6 or control plasmid. For (H), n = 6 independent biological replicates; for the others, n = 3 independent biological replicates. Data are presented as mean ± SD; ∗ P < 0.05, ∗∗ P < 0.01; one-way ANOVA for statistics.

Techniques: Ubiquitin Proteomics, Co-Immunoprecipitation Assay, Transfection, Staining, Control, Plasmid Preparation

Daclatasvir enhances PLIN2 binding, ubiquitination, and interaction with MARCH6 via key residue mutations. A: Molecular docking model demonstrating the binding site (Ser229 and Glu241) of daclatasvir within the active pocket of PLIN2 protein. The upper left box is a zoomed-in display of the lower right box. Red and blue arrows point to daclatasvir and PLIN2 protein, respectively. B: Schematic representation of PLIN2 with mutations in the daclatasvir binding site (PLIN2-mut). PAT domain, Perilipin-ADRP-Tip47 domain; LD, lipid droplet; S, serine; E, glutamate; A, alanine. C: CoIP assay of the interaction between MARCH6 and PLIN2-mut by immunoprecipitating HA-tagged PLIN2-mut in HEK 293T cells treated with DMSO or daclatasvir (10 μM) for 12 h following transfection with the indicated plasmids. D: Ubiquitination levels of PLIN2-mut in HEK 293T cells transfected with HA-tagged PLIN2-mut, and Myc-tagged Ub-K11O, treated with DMSO or daclatasvir (10 μM) in the presence of MG132 (10 μM). E: Ubiquitination levels of PLIN2-mut in HEK 293T cells transfected with HA-tagged PLIN2-mut, Flag-tagged MARCH6, and Myc-tagged Ub-K11O, treated with DMSO or daclatasvir (10 μM) in the presence of MG132 (10 μM). All experiments were performed in three independent biological replicates.

Journal: Journal of Lipid Research

Article Title: Hepatoprotective drug screening identifies daclatasvir, a promising therapeutic candidate for MASLD by targeting PLIN2

doi: 10.1016/j.jlr.2025.100835

Figure Lengend Snippet: Daclatasvir enhances PLIN2 binding, ubiquitination, and interaction with MARCH6 via key residue mutations. A: Molecular docking model demonstrating the binding site (Ser229 and Glu241) of daclatasvir within the active pocket of PLIN2 protein. The upper left box is a zoomed-in display of the lower right box. Red and blue arrows point to daclatasvir and PLIN2 protein, respectively. B: Schematic representation of PLIN2 with mutations in the daclatasvir binding site (PLIN2-mut). PAT domain, Perilipin-ADRP-Tip47 domain; LD, lipid droplet; S, serine; E, glutamate; A, alanine. C: CoIP assay of the interaction between MARCH6 and PLIN2-mut by immunoprecipitating HA-tagged PLIN2-mut in HEK 293T cells treated with DMSO or daclatasvir (10 μM) for 12 h following transfection with the indicated plasmids. D: Ubiquitination levels of PLIN2-mut in HEK 293T cells transfected with HA-tagged PLIN2-mut, and Myc-tagged Ub-K11O, treated with DMSO or daclatasvir (10 μM) in the presence of MG132 (10 μM). E: Ubiquitination levels of PLIN2-mut in HEK 293T cells transfected with HA-tagged PLIN2-mut, Flag-tagged MARCH6, and Myc-tagged Ub-K11O, treated with DMSO or daclatasvir (10 μM) in the presence of MG132 (10 μM). All experiments were performed in three independent biological replicates.

Techniques: Binding Assay, Ubiquitin Proteomics, Residue, Co-Immunoprecipitation Assay, Transfection

Daclatasvir reduces lipid accumulation by targeting PLIN2 and enhancing lipid metabolism pathways. A: PLIN2 protein levels in HuH-7 cells lentiviral infected with either control shRNA or sh PLIN2 . Quantitative data are shown on the bottom, using β-ACTIN as the loading control. B: Representative Nile Red staining images of sh PLIN2 HuH-7 cell lines with the reintroduction of PLIN2 or PLIN2 -mut plasmids, and treated with DMSO or daclatasvir (10 μM) for 12 h following 18 h of PA/OA (0.25 mM/0.5 mM) stimulation. Scale bar, 25 μm. C: The mRNA levels of lipolysis and fatty acid oxidation-related genes, including ATGL , MAGL , ACOX1 , CPT1α , and PPARα , in sh PLIN2 HuH-7 cell lines reintroduced with PLIN2 or PLIN2 -mut plasmids, treated with daclatasvir or DMSO for 12 h in presence of PA (0.5 mM) stimulation. D: Schematic representation illustrating that daclatasvir (DCV) binds to wild type (WT) PLIN2 in the cytoplasm, which facilitates MARCH6-mediated K11 ubiquitination of PLIN2. This process increases the access of PLIN2 to the ubiquitin (Ub)-proteasome system (UPS) and degradation, causes PLIN2 less bound to LD. It diminishes the protective effect of LD-anchored PLIN2, reducing the stability of LD and enhancing lipolysis, resulting in a reduction in lipid accumulation. Because of the non-interaction with daclatasvir, mutant (Mut) PLIN2 get less access to the UPS and anchors more into LD, thus keeping the stability of LD and inhibits lipolysis. For (A, B), n = 3 independent biological replicates; for (C), n = 6 independent biological replicates. Data are presented as mean ± SD; ∗ P < 0.05, ∗∗ P < 0.01, n.s., not significant; Student's t test in (A), and one-way ANOVA in (B, C).

Journal: Journal of Lipid Research

Article Title: Hepatoprotective drug screening identifies daclatasvir, a promising therapeutic candidate for MASLD by targeting PLIN2

doi: 10.1016/j.jlr.2025.100835

Figure Lengend Snippet: Daclatasvir reduces lipid accumulation by targeting PLIN2 and enhancing lipid metabolism pathways. A: PLIN2 protein levels in HuH-7 cells lentiviral infected with either control shRNA or sh PLIN2 . Quantitative data are shown on the bottom, using β-ACTIN as the loading control. B: Representative Nile Red staining images of sh PLIN2 HuH-7 cell lines with the reintroduction of PLIN2 or PLIN2 -mut plasmids, and treated with DMSO or daclatasvir (10 μM) for 12 h following 18 h of PA/OA (0.25 mM/0.5 mM) stimulation. Scale bar, 25 μm. C: The mRNA levels of lipolysis and fatty acid oxidation-related genes, including ATGL , MAGL , ACOX1 , CPT1α , and PPARα , in sh PLIN2 HuH-7 cell lines reintroduced with PLIN2 or PLIN2 -mut plasmids, treated with daclatasvir or DMSO for 12 h in presence of PA (0.5 mM) stimulation. D: Schematic representation illustrating that daclatasvir (DCV) binds to wild type (WT) PLIN2 in the cytoplasm, which facilitates MARCH6-mediated K11 ubiquitination of PLIN2. This process increases the access of PLIN2 to the ubiquitin (Ub)-proteasome system (UPS) and degradation, causes PLIN2 less bound to LD. It diminishes the protective effect of LD-anchored PLIN2, reducing the stability of LD and enhancing lipolysis, resulting in a reduction in lipid accumulation. Because of the non-interaction with daclatasvir, mutant (Mut) PLIN2 get less access to the UPS and anchors more into LD, thus keeping the stability of LD and inhibits lipolysis. For (A, B), n = 3 independent biological replicates; for (C), n = 6 independent biological replicates. Data are presented as mean ± SD; ∗ P < 0.05, ∗∗ P < 0.01, n.s., not significant; Student's t test in (A), and one-way ANOVA in (B, C).

Techniques: Infection, Control, shRNA, Staining, Ubiquitin Proteomics, Mutagenesis

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