do264  (MedChemExpress)

Journal: iScience

Article Title: ABHD12 contributes to tumorigenesis and sorafenib resistance by preventing ferroptosis in hepatocellular carcinoma

doi: 10.1016/j.isci.2023.108340

Figure Lengend Snippet: ABHD12 inhibits sorafenib-induced ferroptosis in HepG2 cells Different HepG2 cell lines were used in the following experiments: Control-OE, cells treated with empty viral vector; WT, wild-type cells; ABHD12-OE , cells overexpressing ABHD12 ; WT-Control, wild-type cells treated with vehicle; ABHD12-KO , cells from which ABHD12 was deleted; DO264, cells treated with ABHD12 inhibitor (DO264). Cells were treated with or without 10 μM sorafenib (Srf) for 24 h. (A) The upper panel shows western blot analysis of GPX4 protein; the lower panel shows the corresponding densitometry. GAPDH was used as an internal control. Data are presented as mean ± SD (n = 3 independent experiments). ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001. (B) Confocal images of lipid reactive oxygen species (ROS), as detected by the fluorescent probe C11-BODIPY 581/591 . Scale bar, 5 μm. (C) Representative confocal images of intracellular ROS levels using the fluorescent probe DCFH-DA. Scale bar, 5 μm. (D) IC 50 values for sorafenib against different cell populations. Results are from three independent experiments. (E) Surface plots for higher order drug combinations. Sorafenib and DO264 act synergistically on HepG2 cells. Sorafenib and DO264 at the indicated concentrations were used to treat cells for 24 h, and cell viability was assessed by MTT assay. ZIP Synergy scores were calculated using Synergyfinder software. Scores > 0 indicated synergism, and scores > 10 were considered strong synergistic. The gradation of the red regions indicates the intensity of synergism. Results are from three independent experiments. (F) Quantification of cell viability as assessed by MTT assay. (G) Representative images of colony formation assay after cells were treated for 2 weeks with sorafenib at 0, 7, or 14 μM in the presence or absence of l0 μM ferrostatin-1 (Fer-1). Results are from three independent experiments. Abbreviations: OE, overexpressing; WT, wild-type; KO, knock out; Srf, sorafenib; ROS, reactive oxygen species; Fer-1, ferrostatin-1; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; GPX4, antioxidant enzyme glutathione peroxidase 4.

Article Snippet: When tumors were palpable, sorafenib (S125098; Aladdin, Shanghai, China) was applied at 20 mg/kg via gavage administered daily for 3 weeks, two groups of animals also received 30 mg/kg DO264 (2301866-59-9, MCE) during the same period.

Techniques: Control, Plasmid Preparation, Western Blot, MTT Assay, Software, Colony Assay, Knock-Out

Journal: iScience

Article Title: ABHD12 contributes to tumorigenesis and sorafenib resistance by preventing ferroptosis in hepatocellular carcinoma

doi: 10.1016/j.isci.2023.108340

Figure Lengend Snippet: ABHD12 contributes to tumorigenesis and sorafenib resistance in liver cancer cells (A) Schematic of the experimental design. Five HepG2 cells lines were used in the following experiments: WT, wild-type cells; ABHD12-OE , cells overexpressing ABHD12 ; WT-Control, cells treated with vehicle; ABHD12-KO , cells from which ABHD12 was deleted; DO264, cells treated with ABHD12 inhibitor (DO264). Cells were implanted into nude mice, which were then treated with 20 mg/kg sorafenib (Srf), 30 mg/kg DO264, or DMSO vehicle. (B) Photographs of tumor weights at sacrifice. (C and D) Tumor sizes were measured once a week. (E) Tumor weight. Data are shown as mean ± SD. Statistical significance was calculated using two-way ANOVA. ∗∗p < 0.01, ∗∗∗p < 0.001. (F) Western blot analysis of GPX4 protein in tumor. GAPDH was used as an internal control. (G) A model for how ABHD12 may contribute to tumorigenesis and sorafenib resistance of HepG2 cells. Abbreviations: OE, overexpressing; WT, wild-type; KO, knock out; LIHC, liver hepatocellular carcinoma; Fer, ferrostatin-1; GPX4, antioxidant enzyme glutathione peroxidase 4.

Article Snippet: When tumors were palpable, sorafenib (S125098; Aladdin, Shanghai, China) was applied at 20 mg/kg via gavage administered daily for 3 weeks, two groups of animals also received 30 mg/kg DO264 (2301866-59-9, MCE) during the same period.

Techniques: Control, Western Blot, Knock-Out

Journal: iScience

Article Title: ABHD12 contributes to tumorigenesis and sorafenib resistance by preventing ferroptosis in hepatocellular carcinoma

doi: 10.1016/j.isci.2023.108340

Figure Lengend Snippet:

Article Snippet: When tumors were palpable, sorafenib (S125098; Aladdin, Shanghai, China) was applied at 20 mg/kg via gavage administered daily for 3 weeks, two groups of animals also received 30 mg/kg DO264 (2301866-59-9, MCE) during the same period.

Techniques: Virus, Recombinant, Expressing, Plasmid Preparation, Software

Journal: bioRxiv

Article Title: Spatially resolved mapping of monoacylglycerol lipase activity in the brain

doi: 10.1101/2025.07.08.663730

Figure Lengend Snippet: a, Mechanism of MAGL labeling by hexafluoroisopropanol-carbamates. b, Structures of the three probes synthesized in this study. The three probes bear the same MAGL targeting element, but are equipped with fluorophores with different excitation/emission wavelengths: LEI-463-BDP-FL , 493/503 nm; LEI-463-BDP-TMR , 542/574 nm, LEI-463-Cy5 , 646/662 nm. c, Dose response labeling of MAGL in mouse brain proteome by the indicated LEI-463 probes, and quantification of IC 50 and EC 50 values (N=3) . d, Absence of MAGL labeling after incubation with probe (100 nM, 30 min, RT) by either pre-incubation with ABX-1431 (10 µM) or in lysates from Mgll /- mice (indicated as KO). e-f, Mouse brain lysates ( e ) or human cortical lysates ( f ) were incubated with the indicated inhibitors prior to treatment with the LEI-463 probes (100 nM). Abbreviations: ABX, ABX-1431; LEI, LEI-515; KT, KT182; DO, DO264.

Article Snippet: KT182 was purchased from Merck (SML1248) and DO264 was purchased from Focus Biomolecules (10-3712).

Techniques: Labeling, Synthesized, Incubation

Journal: iScience

Article Title: Suppressing phagocyte activation by overexpressing the phosphatidylserine lipase ABHD12 preserves sarmopathic nerves

doi: 10.1016/j.isci.2025.112626

Figure Lengend Snippet: SARM1 activation triggers PS dysregulation in stressed-but-viable axons (A and B) Representative (A) brightfield and (B) Annexin V fluorescent images of blebs on dKO axons transduced with SARM1. Insets depict magnified representative images of axonal blebs (arrows) and Annexin V signal on sarmopathic axons (dKO + SARM1). (C) Quantification of Annexin V intensity on axonal blebs from wild-type, dKO, and dKO + SARM1 neurons ( n = 16). (D) Quantification of relative lyso-PS levels as measured by LC-MS/MS after treatment of wild-type or SARM1 KO neurons with DMSO (control) or 50μM Vacor at 0, 2, and 4hrs. (E) Quantification of relative lyso-PS levels as measured by LC-MS/MS after 24h treatment of wild-type neurons with increasing doses of the ABHD12 inhibitor DO264 ( n = 3). (F) Quantification of NAD+ levels after 48h treatment of wild-type neurons with 100nM DO264 ( n = 3). (G) Quantification of ATP levels after 48h treatment of wild-type neurons with 100nM DO264 ( n = 3). (H) Quantification of cADPR levels after 48h treatment of wild-type neurons with 100nM DO264 ( n = 3). All data are presented as mean ± SEM. Statistical significance determined by Student’s unpaired t test or one-way ANOVA with multiple comparisons. ns: not significant, ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001.

Article Snippet: DO264 , Cayman Chemical Company , 29402.

Techniques: Activation Assay, Transduction, Liquid Chromatography with Mass Spectroscopy, Control

Journal: iScience

Article Title: ABHD12 contributes to tumorigenesis and sorafenib resistance by preventing ferroptosis in hepatocellular carcinoma

doi: 10.1016/j.isci.2023.108340

Figure Lengend Snippet: ABHD12 is overexpressed in liver cancer (A) Heatmap of TCGA samples. (B) Genetic alterations and expression of ABHD genes in LIHC. (C) Summary of alterations in differentially expressed ABHD s in LIHC. (D) Comparison of ABHD expression between various cancers and normal tissue. (E) Overall survival of LIHC patients, stratified by ABHD12 expression. (F) Disease-free survival of LIHC patients, stratified by ABHD12 expression. (G) Comparison of ABHD12 expression between LIHC and normal tissues in the TCGA database. ∗∗∗p < 0.001, unpaired t test. (H) Fold changes in ABHD12 gene expression in various cancers. ∗p < 0.05. Abbreviations: ABHD, α/β-hydrolase domain-containing; LIHC, liver hepatocellular carcinoma; BLCA, bladder urothelial carcinoma; BRCA, breast invasive carcinoma; CESC, cervical squamous cell carcinoma and endocervical adenocarcinoma; CHOL, cholangiocarcinoma; COAD, colon adenocarcinoma; DLBC, lymphoid neoplasm diffuse large B cell lymphoma; ESCA, esophageal carcinoma; HNSC, head and neck squamous cell carcinoma; KIRC, kidney renal clear cell carcinoma; LAML, acute myeloid leukemia; LUAD, lung adenocarcinoma; OV, ovarian serous cystadenocarcinoma; THCA thyroid carcinoma; UCEC, uterine corpus endometrial carcinoma. N, normal; T, tumor.

Article Snippet: 4-week-old male BALB/c nude mice placed in five groups (n = 3) and implanted with one of the following HepG2 cell lines: Parental, wild-type cells; ABHD12-OE , cells overexpressing ABHD12 ; Parental-Control, wild-type cells treated with vehicle; ABHD12-KO , cells from which ABHD12 was deleted; DO264, cells treated with ABHD12 inhibitor (DO264) (2301866-59-9, MCE, USA).

Techniques: Expressing, Comparison

Journal: iScience

Article Title: ABHD12 contributes to tumorigenesis and sorafenib resistance by preventing ferroptosis in hepatocellular carcinoma

doi: 10.1016/j.isci.2023.108340

Figure Lengend Snippet: Expression of ABHD12 in liver cancer tissues Tumor and adjacent non-tumor tissues were taken from LIHC patients, and ABHD12 protein levels were quantified. (A) Immunohistochemistry of ABHD12 in cancerous and adjacent non-tumor tissues. H&E: scale bar, 400 μm. Enlarged image: scale bar, 100 μm. (B) Quantitation of ABHD12 expression based on immunohistochemistry (patients, n = 58). (C) Distribution of ABHD12 levels of 58 patients based on immunohistochemistry. (D) Quantitative real-time PCR of ABHD12 mRNA in LIHC tissues and adjacent non-tumor tissues (patients, n = 12). ACTIN was used as an internal control. ∗∗p < 0.01, ∗∗∗p < 0.001. Abbreviations: H&E, hematoxylin and eosin staining.

Article Snippet: 4-week-old male BALB/c nude mice placed in five groups (n = 3) and implanted with one of the following HepG2 cell lines: Parental, wild-type cells; ABHD12-OE , cells overexpressing ABHD12 ; Parental-Control, wild-type cells treated with vehicle; ABHD12-KO , cells from which ABHD12 was deleted; DO264, cells treated with ABHD12 inhibitor (DO264) (2301866-59-9, MCE, USA).

Techniques: Expressing, Immunohistochemistry, Quantitation Assay, Real-time Polymerase Chain Reaction, Control, Staining

Journal: iScience

Article Title: ABHD12 contributes to tumorigenesis and sorafenib resistance by preventing ferroptosis in hepatocellular carcinoma

doi: 10.1016/j.isci.2023.108340

Figure Lengend Snippet: ABHD12 is correlated with different biological processes in LIHC (A and B) The role of ABHD12 in biological processes was investigated by gene set enrichment analysis of the TCGA dataset. ABHD12 expression showed enrichment of genes involved in the cell cycle and mismatch repair. (C and D) Spearman’s correlation analysis identified associations between the level of ABHD12 mRNA and enrichment of genes in the following biological pathways in LIHC: (C) cell cycle ( CDK1 and CDK7 ); (D) mismatch repair ( MLH3 and MSH3 ).(E) The role of ABHD12 in biological processes was investigated by gene set enrichment analysis of the TCGA dataset. ABHD12 expression showed enrichment of genes involved in fatty acid metabolism. (F and G) Spearman’s correlation analysis identified associations between the level of ABHD12 mRNA and enrichment of genes in the following biological pathways in LIHC: (F) positive regulation of ferroptosis ( p53 ); (G) negative regulation of ferroptosis ( GPX4 ) . Abbreviations: CDK1 , cyclin-dependent kinase 1; CDK7 , cyclin-dependent kinase 7; MLH3 , mutL homolog 3; MSH3 , mutS homolog 3; GPX4 , glutathione peroxidase 4.

Article Snippet: 4-week-old male BALB/c nude mice placed in five groups (n = 3) and implanted with one of the following HepG2 cell lines: Parental, wild-type cells; ABHD12-OE , cells overexpressing ABHD12 ; Parental-Control, wild-type cells treated with vehicle; ABHD12-KO , cells from which ABHD12 was deleted; DO264, cells treated with ABHD12 inhibitor (DO264) (2301866-59-9, MCE, USA).

Techniques: Expressing

Journal: iScience

Article Title: ABHD12 contributes to tumorigenesis and sorafenib resistance by preventing ferroptosis in hepatocellular carcinoma

doi: 10.1016/j.isci.2023.108340

Figure Lengend Snippet: ABHD12 promotes proliferation and migration of HepG2 cells Four HepG2 cell lines were used in the following experiments: Control-OE, cells treated with empty viral vector; ABHD12-OE , cells overexpressing ABHD12 ; WT, wild-type cells; ABHD12-KO , cells from which ABHD12 was deleted. (A) Western blot analysis of ABHD12 expression in four HepG2 cell lines; GAPDH was used as an internal control. (B and C) Representative images and quantification of (B) colony formation assays and (C) migration assays. Scale bar, 50 μm. (D and E) Representative images of wound healing experiments before and after injury and quantification of migration within 24 h. Scale bar, 100 μm. (F) Cell proliferation as assessed by MTT assay. (G and H) Fluorescence micrographs of EdU incorporation into cellular DNA and quantification of EdU + cells. Scale bar, 50 μm. All data are presented as the mean ± SD from three independent experiments. ∗p < 0.05, ∗∗p < 0.01, and ∗∗∗p < 0.001 versus control group. Abbreviations: OE, overexpressing; KO, knock out.

Article Snippet: 4-week-old male BALB/c nude mice placed in five groups (n = 3) and implanted with one of the following HepG2 cell lines: Parental, wild-type cells; ABHD12-OE , cells overexpressing ABHD12 ; Parental-Control, wild-type cells treated with vehicle; ABHD12-KO , cells from which ABHD12 was deleted; DO264, cells treated with ABHD12 inhibitor (DO264) (2301866-59-9, MCE, USA).

Techniques: Migration, Control, Plasmid Preparation, Western Blot, Expressing, MTT Assay, Fluorescence, Knock-Out

Journal: iScience

Article Title: ABHD12 contributes to tumorigenesis and sorafenib resistance by preventing ferroptosis in hepatocellular carcinoma

doi: 10.1016/j.isci.2023.108340

Figure Lengend Snippet: ABHD12 inhibits sorafenib-induced ferroptosis in HepG2 cells Different HepG2 cell lines were used in the following experiments: Control-OE, cells treated with empty viral vector; WT, wild-type cells; ABHD12-OE , cells overexpressing ABHD12 ; WT-Control, wild-type cells treated with vehicle; ABHD12-KO , cells from which ABHD12 was deleted; DO264, cells treated with ABHD12 inhibitor (DO264). Cells were treated with or without 10 μM sorafenib (Srf) for 24 h. (A) The upper panel shows western blot analysis of GPX4 protein; the lower panel shows the corresponding densitometry. GAPDH was used as an internal control. Data are presented as mean ± SD (n = 3 independent experiments). ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001. (B) Confocal images of lipid reactive oxygen species (ROS), as detected by the fluorescent probe C11-BODIPY 581/591 . Scale bar, 5 μm. (C) Representative confocal images of intracellular ROS levels using the fluorescent probe DCFH-DA. Scale bar, 5 μm. (D) IC 50 values for sorafenib against different cell populations. Results are from three independent experiments. (E) Surface plots for higher order drug combinations. Sorafenib and DO264 act synergistically on HepG2 cells. Sorafenib and DO264 at the indicated concentrations were used to treat cells for 24 h, and cell viability was assessed by MTT assay. ZIP Synergy scores were calculated using Synergyfinder software. Scores > 0 indicated synergism, and scores > 10 were considered strong synergistic. The gradation of the red regions indicates the intensity of synergism. Results are from three independent experiments. (F) Quantification of cell viability as assessed by MTT assay. (G) Representative images of colony formation assay after cells were treated for 2 weeks with sorafenib at 0, 7, or 14 μM in the presence or absence of l0 μM ferrostatin-1 (Fer-1). Results are from three independent experiments. Abbreviations: OE, overexpressing; WT, wild-type; KO, knock out; Srf, sorafenib; ROS, reactive oxygen species; Fer-1, ferrostatin-1; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; GPX4, antioxidant enzyme glutathione peroxidase 4.

Article Snippet: 4-week-old male BALB/c nude mice placed in five groups (n = 3) and implanted with one of the following HepG2 cell lines: Parental, wild-type cells; ABHD12-OE , cells overexpressing ABHD12 ; Parental-Control, wild-type cells treated with vehicle; ABHD12-KO , cells from which ABHD12 was deleted; DO264, cells treated with ABHD12 inhibitor (DO264) (2301866-59-9, MCE, USA).

Techniques: Control, Plasmid Preparation, Western Blot, MTT Assay, Software, Colony Assay, Knock-Out

Journal: iScience

Article Title: ABHD12 contributes to tumorigenesis and sorafenib resistance by preventing ferroptosis in hepatocellular carcinoma

doi: 10.1016/j.isci.2023.108340

Figure Lengend Snippet: ABHD12 contributes to tumorigenesis and sorafenib resistance in liver cancer cells (A) Schematic of the experimental design. Five HepG2 cells lines were used in the following experiments: WT, wild-type cells; ABHD12-OE , cells overexpressing ABHD12 ; WT-Control, cells treated with vehicle; ABHD12-KO , cells from which ABHD12 was deleted; DO264, cells treated with ABHD12 inhibitor (DO264). Cells were implanted into nude mice, which were then treated with 20 mg/kg sorafenib (Srf), 30 mg/kg DO264, or DMSO vehicle. (B) Photographs of tumor weights at sacrifice. (C and D) Tumor sizes were measured once a week. (E) Tumor weight. Data are shown as mean ± SD. Statistical significance was calculated using two-way ANOVA. ∗∗p < 0.01, ∗∗∗p < 0.001. (F) Western blot analysis of GPX4 protein in tumor. GAPDH was used as an internal control. (G) A model for how ABHD12 may contribute to tumorigenesis and sorafenib resistance of HepG2 cells. Abbreviations: OE, overexpressing; WT, wild-type; KO, knock out; LIHC, liver hepatocellular carcinoma; Fer, ferrostatin-1; GPX4, antioxidant enzyme glutathione peroxidase 4.

Article Snippet: 4-week-old male BALB/c nude mice placed in five groups (n = 3) and implanted with one of the following HepG2 cell lines: Parental, wild-type cells; ABHD12-OE , cells overexpressing ABHD12 ; Parental-Control, wild-type cells treated with vehicle; ABHD12-KO , cells from which ABHD12 was deleted; DO264, cells treated with ABHD12 inhibitor (DO264) (2301866-59-9, MCE, USA).

Techniques: Control, Western Blot, Knock-Out

Journal: iScience

Article Title: ABHD12 contributes to tumorigenesis and sorafenib resistance by preventing ferroptosis in hepatocellular carcinoma

doi: 10.1016/j.isci.2023.108340

Figure Lengend Snippet:

Article Snippet: 4-week-old male BALB/c nude mice placed in five groups (n = 3) and implanted with one of the following HepG2 cell lines: Parental, wild-type cells; ABHD12-OE , cells overexpressing ABHD12 ; Parental-Control, wild-type cells treated with vehicle; ABHD12-KO , cells from which ABHD12 was deleted; DO264, cells treated with ABHD12 inhibitor (DO264) (2301866-59-9, MCE, USA).

Techniques: Virus, Recombinant, Expressing, Plasmid Preparation, Software