diallyl trisulfide  (Tokyo Chemical Industry)

Journal: Journal of Cancer

Article Title: Diallyl Trisulfide Enhances Doxorubicin Chemosensitivity by Inhibiting the Warburg Effect and Inducing Apoptosis in Breast Cancer Cells

doi: 10.7150/jca.113578

Figure Lengend Snippet: Diallyl trisulfide demonstrates selective cytotoxicity in normal and cancer cells. (A) Normal cell lines (MCF10, Clone 9, Beas-2B, and AC16) were treated with increasing concentrations of DATS for 24 hours followed by MTT cell viability assay. (B) Cancer cell lines (MCF7, MDA-MB-231, A549, LoVo, and HA22T) were exposed to the same DATS treatment as in (A) , followed by MTT cell viability assay. These results are expressed as a percentage of viable cells from treated groups compared with control. All experiments were conducted in triplicate, ns=not significant, **P < 0.001 and ***P < 0.0001.

Article Snippet: Diallyl trisulfide (CAS No. 250-87-5) was procured from MedChemExpress (USA).

Techniques: Viability Assay, Control

Journal: Journal of Cancer

Article Title: Diallyl Trisulfide Enhances Doxorubicin Chemosensitivity by Inhibiting the Warburg Effect and Inducing Apoptosis in Breast Cancer Cells

doi: 10.7150/jca.113578

Figure Lengend Snippet: Diallyl trisulfide synergistically enhances doxorubicin cytotoxicity in breast cancer cells. (A and B) Breast cancer cells—(A) MCF7 and (B) MDA-MB-231—were treated with the indicated concentrations of DOX for 24 and 48 hours, followed by MTT cell viability assay. (C and D) Breast cancer cells — (C) MCF7 and (D) MDA-MB-231— were treated with the indicated concentrations of DATS for 24 and 48 hours, followed by the MTT cell viability assay. (E and F) Breast cancer cells— (E) MCF7 and (F) MDA-MB-231—were pre-treated with Dox (0.5 and 1 µM) for 24 hours, followed by the addition of DATS (50 and 100 µM) for a combined treatment period of 48 hours, followed by MTT cell viability assay. (G and H) Combination index (CI) calculation in (G) MCF7 and (H) MDA-MB-231 cells. The CI reflects the degree of drug-drug interactions; in this study, CIs <0.9 indicated synergism, CIs ranging from 0.9 to 1.1 indicated additive effects, and CIs >1.1 indicated antagonistic effects. These results are expressed as a percentage of viable cells from treated groups compared with control cells, ns= not significant, *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: Diallyl trisulfide (CAS No. 250-87-5) was procured from MedChemExpress (USA).

Techniques: Viability Assay, Control

Journal: Journal of Cancer

Article Title: Diallyl Trisulfide Enhances Doxorubicin Chemosensitivity by Inhibiting the Warburg Effect and Inducing Apoptosis in Breast Cancer Cells

doi: 10.7150/jca.113578

Figure Lengend Snippet: Diallyl trisulfide and doxorubicin combination inhibits glucose metabolism in breast cancer cells. (A ) MCF7 and MDA-MB-231 breast cancer cells were treated with DOX (0.5 μM, 1 μM), DATS (50 μM, 100 μM), and their combination, followed by 2-NBDG glucose uptake flow cytometry analysis. (B) The same treatments as in (A) followed by western blot analysis of key glycolytic regulators: GLUT1, PDH, LDHA, and HIF-1α in MCF7 cancer cells. (C) Similar treatment as in A and immunoblotting as in B for MDA-MB-231 cells (D) Similar treatments as in (A) followed by a lactate production colorimetric assay. (E) Lactate production colorimetric assay in MDA-MB-231 cells exposed to LDHA-specific siRNA (50 nM) with or without Doxorubicin (0.5 and 1 µM). (F) Western blot analysis of LDHA, GLUT1 and HIF1α in MDA-MB-231 cells treated with LDHA-siRNA (50 nM) with or without DOX (1 µM). B-Actin is loading control. Data in panels (A), (D), and (E) are expressed as the mean ± SD of treated groups compared to the control, ns=not significant, *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: Diallyl trisulfide (CAS No. 250-87-5) was procured from MedChemExpress (USA).

Techniques: Flow Cytometry, Western Blot, Colorimetric Assay, Control

Journal: Journal of Cancer

Article Title: Diallyl Trisulfide Enhances Doxorubicin Chemosensitivity by Inhibiting the Warburg Effect and Inducing Apoptosis in Breast Cancer Cells

doi: 10.7150/jca.113578

Figure Lengend Snippet: Diallyl trisulfide and doxorubicin combination induces apoptosis in breast cancer cells. (A) MCF7 cancer cells were treated with a combination of DOX (0.5 and 1 μM) and DATS (50 and 100 μM), followed by western blot analysis of the indicated apoptosis markers. (B) MDA-MB-231 cancer cells were treated similarly to (A), followed by western blot analysis. β-Actin was used as the loading control. (C) MCF7 cancer cells were treated similarly to (A), followed by TUNEL staining. (D) MDA-MB-231 cancer cells were treated similarly to (A), followed by TUNEL staining. (E) Annexin V-FITC/PI staining and flow cytometry analysis in MCF7 and MDA-MB-231 cells treated similarly to (A). Data in panels (C), (D), and (E) are expressed as the mean ± SD of treated groups compared to the control, ns=not significant, *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: Diallyl trisulfide (CAS No. 250-87-5) was procured from MedChemExpress (USA).

Techniques: Western Blot, Control, TUNEL Assay, Staining, Flow Cytometry

Journal: Journal of Cancer

Article Title: Diallyl Trisulfide Enhances Doxorubicin Chemosensitivity by Inhibiting the Warburg Effect and Inducing Apoptosis in Breast Cancer Cells

doi: 10.7150/jca.113578

Figure Lengend Snippet: Diallyl trisulfide and doxorubicin combination inhibits MDA-MB-231 breast cancer cell growth in vivo . (A) Combination treatment in vivo experimental design. (B) Monitoring of mouse body weight during the treatment period. (C) Tumor volume monitoring during treatment period. (D) Kaplan-Meier survival analysis from tumor initiation to experiment end point. (E) Bioluminescence imaging of tumor-bearing mice at the end of the experiment. (F) Gross tumor size and weight from each group at the end of the experiment. (G) Western blot analysis of apoptotic markers (BAX, Bcl-xL) and metabolic markers (LDHA) in tumor tissue lysates, B-Actin is loading control. (H) H&E staining (upper panels) and LDHA immunohistochemistry (lower panels) in tumor sections. Representative images are shown, scale bar = 50 nm. Data are expressed as the mean ± SD of treated groups compared to the control, ns=not significant, **P < 0.01, ***P < 0.001.

Article Snippet: Diallyl trisulfide (CAS No. 250-87-5) was procured from MedChemExpress (USA).

Techniques: In Vivo, Imaging, Western Blot, Control, Staining, Immunohistochemistry

Journal: Frontiers in Pharmacology

Article Title: Hydrogen sulfide and ferroptosis inhibition underlies the dietary restriction-induced protection against cyclophosphamide cystitis

doi: 10.3389/fphar.2025.1562852

Figure Lengend Snippet: Diallyl trisulfide (DATS) supplementation protects against CYP-induced cystitis. (A) Effect of DATS on bladder morphology. Mice were given 60 mg/kg DATS for 3 days via gavage. Afterward, the mice were i.p. injected with 150 mg/kg CYP. The bladder changes at 12 h were examined. Note the obvious congestion, enlargement, and edema in bladders in CYP group, but a significant improvement in CYP-DATS group. (B) Effect of DATS on bladder weight-to-body weight ratio. Data shown are mean ± S.E. (n = 5; * P < 0.05, ** P < 0.01). (C,D) Effect of DATS on bladder histopathology. Representative histological images of bladder sections from different treatment groups. Scale bar = 100 μm (C) . The thickness of bladder mucosa was analyzed as the percentage of control (D) (mean ± S.E.; n = 5; ** P < 0.01). (E–G) Effect of DATS on CYP-induced bladder oxidative injury. The level of the bladder -SH groups and IgG was determined with a maleimide-labeling assay and Western blot analysis, respectively (E) . Densitometric analysis of the blots was shown in (F,G) and expressed as the percentage of control (mean analysis of the blots was shown in (F,G) and expressed as the percentage of control (mean ± S.E.; n = 3; * P < 0.05, ** P < 0.01).

Article Snippet: Diallyl trisulfide (DATS, #D854967) was obtained from Macklin (Shanghai, China).

Techniques: Injection, Histopathology, Control, Labeling, Western Blot

Journal: Frontiers in Pharmacology

Article Title: Hydrogen sulfide and ferroptosis inhibition underlies the dietary restriction-induced protection against cyclophosphamide cystitis

doi: 10.3389/fphar.2025.1562852

Figure Lengend Snippet: DATS prevents CYP-induced ferroptosis in the bladder. (A) Effect of DATS on bladder MDA production. The extracted bladder proteins were subjected to MDA assay following the manufacturer’s protocol (mean ± S.E.; n = 5; ** P < 0.01). (B–D) Effect of DATS on ferroptosis markers in mouse bladder. Bladder proteins were analyzed with Western blot for SLC7A11, ACSL4, and GAPDH expression (B) . Densitometric analysis of the blots was shown in (C,D) and expressed as the percentage of control (mean ± S.E.; n = 3; ** P < 0.01).

Article Snippet: Diallyl trisulfide (DATS, #D854967) was obtained from Macklin (Shanghai, China).

Techniques: Multiple Displacement Amplification, Western Blot, Expressing, Control

Journal: Frontiers in Pharmacology

Article Title: Hydrogen sulfide and ferroptosis inhibition underlies the dietary restriction-induced protection against cyclophosphamide cystitis

doi: 10.3389/fphar.2025.1562852

Figure Lengend Snippet: NaHS and DATS protect against ACR-induced urothelial cell injury. (A,B) Effect of DATS on H 2 S production capacity in urothelial cells. SV-HUC-1 cells treated with indicated concentrations of DATS were subjected to lead sulfide assay for 24 h (A) . Densitometric quantification of the intensity of the colored circles was performed, and data shown in (B) have been normalized to control (mean ± S.E.; n = 5; ** P < 0.01). (C,D) Effect of NaHS and DATS on ACR-induced urothelial cell death. SV-HUC-1 cells were pretreated with 1 mM NaHS for 15 min or 25 μM DATS for 30 min before exposing to 100 μM ACR for an additional 24 h. The cell viability was determined by Calcein-AM/PI staining (upper) and WST assay (lower). Data shown are mean ± S.E. (n = 5 and 6, respectively; ** P < 0.01). (E,F) Effect of NaHS and DATS on ACR-induced oxidative stress in urothelial cells. SV-HUC-1 cells were pretreated with 1 mM NaHS for 15 min or 25 μM DATS for 30 min before exposing to 75 μM ACR for an additional 4 h. Cell proteins were extracted for Western blot analysis of protein carbonylation and phosphorylated P38MAPK. A quantitative analysis of blots is shown below. Data are presented as mean ± S.E. (n = 3; * P < 0.05, ** P < 0.01).

Article Snippet: Diallyl trisulfide (DATS, #D854967) was obtained from Macklin (Shanghai, China).

Techniques: Control, Staining, WST Assay, Western Blot

Journal: Nutrients

Article Title: The Garlic Compound, Diallyl Trisulfide, Attenuates Benzo[a]Pyrene-Induced Precancerous Effect through Its Antioxidant Effect, AhR Inhibition, and Increased DNA Repair in Human Breast Epithelial Cells

doi: 10.3390/nu16020300

Figure Lengend Snippet: ( A ) The effect of DATS on the viability of MCF-10A cells. MCF-10A cells were treated with 0–100 μM DATS for 24 and 48 h. DATS had no significant effect between 12.5 and 75 μM. Treatment with 100 μM of DATS at 48 h significantly decreased cell viability compared to control. The graph displays all experiments conducted in n = 8 and averaged for three separate experiments. The data were assessed using one-way ANOVA and Dunnett’s multiple comparison test. The average values ± SEM display the results to determine significant differences between the vehicle control and treatments. (* p < 0.05 and ns indicates no significance). ( B ) The effect of B[a]P on viability of MCF-10A cells. MCF-10A cells were treated with 0.01–1 μM B[a]P for 24 and 48 h. Treatment with 1 μM B[a]P and above caused a significant increase in cell viability compared to the control. The graph displays all experiments conducted in n = 8 and averaged for three separate experiments. The data were assessed using one-way ANOVA and Dunnett’s multiple comparison test. The average values ± SEM display the results to determine significant differences between the vehicle control and treatments. (**** p < 0.0001 and ns indicates no significance).

Article Snippet: The diallyl trisulfide (DATS) (purity 99.2%, 200 mM stock) was purchased from LKT Laboratories (St. Paul, MN, USA).

Techniques: Control, Comparison

Journal: Nutrients

Article Title: The Garlic Compound, Diallyl Trisulfide, Attenuates Benzo[a]Pyrene-Induced Precancerous Effect through Its Antioxidant Effect, AhR Inhibition, and Increased DNA Repair in Human Breast Epithelial Cells

doi: 10.3390/nu16020300

Figure Lengend Snippet: Cell proliferation percentage of MCF-10A cells treated with B[a]P and DATS. MCF-10A cells were treated with 1 μM B[a]P only, 40–80 μM DATS only, or 1 μM B[a]P + 40–80 μM CoTx for 12 and 24 h. The graph displays all experiments conducted in n = 8 and averaged for three separate experiments. The data were assessed using one-way ANOVA and Dunnett’s multiple comparison test. The average values ± SEM display the results to determine significant differences between DMSO vehicle, B[a]P, and various treatment groups. (ns indicates no significance, **** p < 0.0001 compared to the control, and #### p < 0.0001 when compared to B[a]P treatment).

Article Snippet: The diallyl trisulfide (DATS) (purity 99.2%, 200 mM stock) was purchased from LKT Laboratories (St. Paul, MN, USA).

Techniques: Comparison, Control

Journal: Nutrients

Article Title: The Garlic Compound, Diallyl Trisulfide, Attenuates Benzo[a]Pyrene-Induced Precancerous Effect through Its Antioxidant Effect, AhR Inhibition, and Increased DNA Repair in Human Breast Epithelial Cells

doi: 10.3390/nu16020300

Figure Lengend Snippet: ( A – F ) Clonogenic formation of MCF-10A cells treated with B[a]P, DATS, and DATS CoTx. ( A ) Effects of B[a]P on colony formation of MCF-10A. ( C ) Effects of DATS on colony formation of MCF-10A cells. ( E ) Effects of 1 μM B[a]P alone, 40 μM DATS alone, and 40 μM CoTx on colony formation of MCF-10A cells. Cells were placed in phenol red-free DMEM supplement with 5% dextran-coated charcoal-treated HS for 24 h before plating. Then 250 cells/well were plated in six-well plates. Seven days later, cells were treated with 0.1% DMSO vehicle control. ( B , D , F ), and the graphs display all experiments conducted in n = 3 and averaged for three separate experiments. The data were assessed using one-way ANOVA and Dunnett’s multiple comparison test. The average values ± SEM display the results to determine significant differences between DMSO vehicle, B[a]P, and various treatment groups. (*** p < 0.001, **** p < 0.0001 compared to the control, and #### p < 0.0001 when compared to B[a]P treatment).

Article Snippet: The diallyl trisulfide (DATS) (purity 99.2%, 200 mM stock) was purchased from LKT Laboratories (St. Paul, MN, USA).

Techniques: Control, Comparison

Journal: Nutrients

Article Title: The Garlic Compound, Diallyl Trisulfide, Attenuates Benzo[a]Pyrene-Induced Precancerous Effect through Its Antioxidant Effect, AhR Inhibition, and Increased DNA Repair in Human Breast Epithelial Cells

doi: 10.3390/nu16020300

Figure Lengend Snippet: DATS inhibition of B[a]P-induced ROS in MCF-10A cells. The cells analyzed for ROS production were treated with B[a]P, DATS, or CoTx for 12 and 24 h. 0.1% Hydrogen peroxide was used as a positive control. The graphs display all experiments conducted in n = 3 and averaged for three separate experiments. The average values ± SEM display the results to determine significant differences between DMSO vehicle, B[a]P, and various treatment groups. (ns indicates no significance, * p < 0.05, ** p < 0.01 compared to the control, and ## p < 0.01 when compared to B[a]P treatment).

Article Snippet: The diallyl trisulfide (DATS) (purity 99.2%, 200 mM stock) was purchased from LKT Laboratories (St. Paul, MN, USA).

Techniques: Inhibition, Positive Control, Control

Journal: Nutrients

Article Title: The Garlic Compound, Diallyl Trisulfide, Attenuates Benzo[a]Pyrene-Induced Precancerous Effect through Its Antioxidant Effect, AhR Inhibition, and Increased DNA Repair in Human Breast Epithelial Cells

doi: 10.3390/nu16020300

Figure Lengend Snippet: DNA damage detection of MCF-10A cells treated with DATS and/or B[a]P. MCF-10A cells were treated with 1 μM B[a]P only or 1 μM B[a]P + 40–80 μM CoTx for 24 h. The graph displays oxidative DNA damage indicated by 8-OHdG in pg. All experiments were conducted in n = 3 and averaged for three separate experiments. The data were assessed using an unpaired t -test. The average values ± SEM display the results to determine significant differences between DMSO vehicle, B[a]P, and various treatment groups. (** p < 0.01, *** p < 0.001, **** p < 0.0001 compared to the control, and #### p < 0.0001 when compared to B[a]P treatment).

Article Snippet: The diallyl trisulfide (DATS) (purity 99.2%, 200 mM stock) was purchased from LKT Laboratories (St. Paul, MN, USA).

Techniques: Control

Journal: Nutrients

Article Title: The Garlic Compound, Diallyl Trisulfide, Attenuates Benzo[a]Pyrene-Induced Precancerous Effect through Its Antioxidant Effect, AhR Inhibition, and Increased DNA Repair in Human Breast Epithelial Cells

doi: 10.3390/nu16020300

Figure Lengend Snippet: DATS and/or B[a]P effect on AhR expression in MCF-10A cells. Protein expression of AhR was measured by densitometry and normalized ( A ). The immunoblots represented the protein expression after 24 h post-treatment for AhR ( B ). The graphs display all experiments conducted in n = 3 and averaged for three separate experiments. The average values ± SEM display the results to determine significant differences between DMSO vehicle, B[a]P, and treatment groups. (ns indicates no significance, *** p < 0.001, **** p < 0.0001 compared to the control, and #### p < 0.0001 when compared to B[a]P treatment).

Article Snippet: The diallyl trisulfide (DATS) (purity 99.2%, 200 mM stock) was purchased from LKT Laboratories (St. Paul, MN, USA).

Techniques: Expressing, Western Blot, Control

Journal: Nutrients

Article Title: The Garlic Compound, Diallyl Trisulfide, Attenuates Benzo[a]Pyrene-Induced Precancerous Effect through Its Antioxidant Effect, AhR Inhibition, and Increased DNA Repair in Human Breast Epithelial Cells

doi: 10.3390/nu16020300

Figure Lengend Snippet: DATS and/or B[a]P effect on HIF-1β expression in MCF-10A. Protein expression of HIF-1β was measured by densitometry and normalized ( A ). The immunoblots represented the protein expression after 24 h post-treatment for HIF-1β ( B ). The graphs display all experiments conducted in n = 3 and averaged for three separate experiments. The average values ± SEM display the results to determine significant differences between DMSO vehicle, B[a]P, and treatment groups. (ns indicates no significance, ** p < 0.01, **** p < 0.0001 compared to the control, and ## p < 0.01, #### p < 0.0001 when compared to B[a]P treatment).

Article Snippet: The diallyl trisulfide (DATS) (purity 99.2%, 200 mM stock) was purchased from LKT Laboratories (St. Paul, MN, USA).

Techniques: Expressing, Western Blot, Control

Journal: Nutrients

Article Title: The Garlic Compound, Diallyl Trisulfide, Attenuates Benzo[a]Pyrene-Induced Precancerous Effect through Its Antioxidant Effect, AhR Inhibition, and Increased DNA Repair in Human Breast Epithelial Cells

doi: 10.3390/nu16020300

Figure Lengend Snippet: DATS and/or B[a]P effect on CYP1A1 expression in MCF-10A cells. Protein expression of CYP1A1 was measured by densitometry and normalized ( A ). The immunoblots represented the protein expression after 24 h post-treatment for CYP1A1 ( B ). The graphs display all experiments conducted in n = 3 and averaged for three separate experiments. The average values ± SEM display the results to determine significant differences between DMSO vehicle, B[a]P, and treatment groups. (ns indicates no significance, **** p < 0.0001 compared to the control and #### p < 0.0001 when compared to B[a]P treatment).

Article Snippet: The diallyl trisulfide (DATS) (purity 99.2%, 200 mM stock) was purchased from LKT Laboratories (St. Paul, MN, USA).

Techniques: Expressing, Western Blot, Control

Journal: Nutrients

Article Title: The Garlic Compound, Diallyl Trisulfide, Attenuates Benzo[a]Pyrene-Induced Precancerous Effect through Its Antioxidant Effect, AhR Inhibition, and Increased DNA Repair in Human Breast Epithelial Cells

doi: 10.3390/nu16020300

Figure Lengend Snippet: DATS and/or B[a]P effect on DNA POLβ expression in MCF-10A. Normalized DNA POLβ protein expression was measured by the ProteinSimple SW Compass software for each of the five samples, one of which is a control, and the others are treated with DMSO, DATS, B[a]P, and CoTx ( A ). Immunoblots from the ProteinSimple SW Compass 6.2.0 software corresponded to the protein expression for the same sample treatments ( B ). The graphs display all experiments conducted in n = 3 and averaged for three separate experiments. The average values ± SEM display the results to determine significant differences between DMSO vehicle, B[a]P, and treatment groups. (ns indicates no significance, ** p < 0.01 compared to the control, and # p < 0.05 when compared to B[a]P treatment).

Article Snippet: The diallyl trisulfide (DATS) (purity 99.2%, 200 mM stock) was purchased from LKT Laboratories (St. Paul, MN, USA).

Techniques: Expressing, Software, Control, Western Blot

Journal: Nutrients

Article Title: The Garlic Compound, Diallyl Trisulfide, Attenuates Benzo[a]Pyrene-Induced Precancerous Effect through Its Antioxidant Effect, AhR Inhibition, and Increased DNA Repair in Human Breast Epithelial Cells

doi: 10.3390/nu16020300

Figure Lengend Snippet: Schematic diagram and graphical abstract for our proposed mechanism of DATS in B[a]P-induced normal breast epithelial MCF-10A cells. The green arrow indicates increase, and the purple arrow indicates decrease. DATS can inhibit clonogenic formation. Whereas DATS CoTx can upregulate DNA repair and DNA POLβ, conversely inhibiting cell proliferation, clonogenic formation, and oxidative stress in the form of ROS. B[a]P can induce oxidative stress, thus upregulating ROS formation, AhR, HIF-1β, and CYP1A1, leading to an increase in DNA damage in the form of DNA single-strand breaks and DNA adducts, increasing cell proliferation and clonogenic formation.

Article Snippet: The diallyl trisulfide (DATS) (purity 99.2%, 200 mM stock) was purchased from LKT Laboratories (St. Paul, MN, USA).

Techniques:

Journal: Cancers

Article Title: Diallyl Trisulfide Induces ROS-Mediated Mitotic Arrest and Apoptosis and Inhibits HNSCC Tumor Growth and Cancer Stemness.

doi: 10.3390/cancers16020378

Figure Lengend Snippet: Figure 1. DATS inhibits cell viability and induces cell cycle arrest in head and neck cancer cells. (A) Structure of DATS. Effect of DATS on Cal33 (B), UMSCC-22A (C) and UMSCC-22B (D). Cells were treated with vehicle (DMSO) alone or 10–40 µM of DATS in a fresh medium. After 24, 48, and 72 h of these treatments, viable cells were counted using trypan blue staining and hemocytometer. DATS treatment caused G2/M phase cell cycle arrest in HNSCC cells. Representative flow histograms depicting cell cycle distribution in UMSCC-22A (E) and UMSCC-22B (F) cell cultures following 8 h treatment with the indicated concentrations of DATS. Quantitative analysis of UMSCC-22A (G) and UMSCC-22B (H) cells in different phases of the cell cycle after treatment with indicated concentrations of DATS. (I) Western blotting for cell cycle regulatory proteins using lysate from UMSCC-22B cells treated with DMSO control and DATS (20, 40 µM) for 24 h. The results shown are mean ± SEM (n = 3). DATS, Diallyl trisulfide, p < 0.05 (*), p < 0.001 (**), p < 0.0001 (***). The original western blot of Figure 1I is in Figure S6.

Article Snippet: Diallyl trisulfide Cancers 2024, 16, 378 3 of 19 (DATS, purity > 98%) was procured from LKT Laboratories (St. Paul, MN, USA), and RNase A was sourced from Promega (Madison, WI, USA).

Techniques: Staining, Western Blot, Control

Journal: Cancers

Article Title: Diallyl Trisulfide Induces ROS-Mediated Mitotic Arrest and Apoptosis and Inhibits HNSCC Tumor Growth and Cancer Stemness.

doi: 10.3390/cancers16020378

Figure Lengend Snippet: Figure 2. DATS treatment caused ROS-mediated mitotic arrest and altered cell cycle regulatory proteins in HNSCC cells. Representative flow histogram depicting Ser-10 phosphorylated histone H3 in UMSCC-22A (A) and UMSCC-22B (C) cells treated for 24 h with DMSO (control) or 40 µM DATS. Quantitation of the percentage of mitotic fraction in UMSCC-22A (B) and UMSCC-22B (D) cells treated with DMSO (control) or 40 µM DATS. DCFDA assay for ROS generation in DATS (40 µM)

Article Snippet: Diallyl trisulfide Cancers 2024, 16, 378 3 of 19 (DATS, purity > 98%) was procured from LKT Laboratories (St. Paul, MN, USA), and RNase A was sourced from Promega (Madison, WI, USA).

Techniques: Control, Quantitation Assay

Journal: Cancers

Article Title: Diallyl Trisulfide Induces ROS-Mediated Mitotic Arrest and Apoptosis and Inhibits HNSCC Tumor Growth and Cancer Stemness.

doi: 10.3390/cancers16020378

Figure Lengend Snippet: Figure 3. DATS-induced ROS-mediated apoptotic cell death attenuated by NAC pretreatment. Cells were treated with either DMSO or different doses of DATS for 24 h. At the end of treatments, cells were harvested and stained with annexin V and PI, and apoptotic cells were analyzed by flow cytometry. Representative histograms of UMSCC-22A (A) and UMSCC-22B (D) cells treated with DMSO (control) or 20 µM and 40 µM DATS for 24 h. Quantified data of (B) percent early and (C) total apoptotic cells of UMSCC-22A cells. Quantified data of (E) percent early and (F) total apoptotic cells of UMSCC-22B. (G) Representative histograms of UMSCC-22B cells treated with DMSO (control) or 40 µM DATS for 24 h in the absence or presence of NAC pretreatment. Quantified data of (H) percent early and (I) total apoptotic cells of UMSCC-22B cells. Data are shown as mean ± SEM of triplicate samples. DATS, Diallyl trisulfide, p < 0.001 (**), p < 0.0001 (***).

Article Snippet: Diallyl trisulfide Cancers 2024, 16, 378 3 of 19 (DATS, purity > 98%) was procured from LKT Laboratories (St. Paul, MN, USA), and RNase A was sourced from Promega (Madison, WI, USA).

Techniques: Staining, Flow Cytometry, Control

Journal: Cancers

Article Title: Diallyl Trisulfide Induces ROS-Mediated Mitotic Arrest and Apoptosis and Inhibits HNSCC Tumor Growth and Cancer Stemness.

doi: 10.3390/cancers16020378

Figure Lengend Snippet: Figure 4. DATS induced ROS-mediated DNA damage and apoptosis in HNSCC cells and altered apoptotic regulatory proteins. (A) Representative images were captured using a fluorescence micro- scope at 200× magnification in UMSCC-22B cells (scale bars = 50 µm). (B) Quantification of comet tail length in UMSCC-22B cells treated with either DMSO control or 20 and 40 µM of DATS. At least 50 cells were used for statistical analysis. (C) Representative images were captured using a fluorescence microscope at 200× magnification in each case in the presence or absence of NAC pre- treatment (scale bars = 50 µm). (D) quantified data of comet tail length. NAC pretreatment attenuated DATS-induced DNA damage. (E) Western blotting for apoptotic regulatory and DNA damage-related proteins using lysate from UMSCC-22B cells treated with DMSO control and DATS (20, 40 µM) for 24 h. (F) Western blotting for mitotic marker, apoptotic regulatory and DNA damage-related proteins using lysate from UMSCC-22B cells pretreated with NAC then with DMSO control and DATS (20, 40 µM) for 24 h. Data are shown as mean ± SEM of triplicate samples. DATS, Diallyl trisulfide, p < 0.0001 (***). The original western blot of Figure 4E,F is in Figure S6.

Article Snippet: Diallyl trisulfide Cancers 2024, 16, 378 3 of 19 (DATS, purity > 98%) was procured from LKT Laboratories (St. Paul, MN, USA), and RNase A was sourced from Promega (Madison, WI, USA).

Techniques: Fluorescence, Control, Microscopy, Western Blot, Marker

Journal: Cancers

Article Title: Diallyl Trisulfide Induces ROS-Mediated Mitotic Arrest and Apoptosis and Inhibits HNSCC Tumor Growth and Cancer Stemness.

doi: 10.3390/cancers16020378

Figure Lengend Snippet: Figure 5. DATS decreased CSC population and stemness-related genes in HNSCC cells. (A) Repre- sentative histograms for CD133high/CD44high fraction of UMSCC-22A cells after 72-h treatment with DMSO or DATS (0–5 µM). (B) Quantitation of CD133high/CD44high fraction of UMSCC-22A cells after DATS treatment. (C) Representative flow histograms for ALDH1 activity in UMSCC-22A cells after 24-h treatment with DMSO or DATS (0–10 µM). The ALDH1 inhibitor DEAB was used as a control. (D) Quantitation of ALDH1 activity of UMSCC-22A cells after DATS treatment. (E) Representative images of spheroids resulting after 14 days of cell seeding and treatment of UMSCC-22A cells with DMSO or DATS (0–10 µM) (magnification ×100, scale bars = 200 µm). (F) Quantitation of number of spheroids. Cells were treated with either DMSO or 20 and 40 µM DATS for 24 h followed by quantitative analysis of stem cell-related genes SOX2 and Oct4 mRNA expression by real-time PCR in UMSCC-22A (G) and UMSCC-22B (H) cells relative to DMSO control. The results shown are relative to the DMSO-treated control (mean ± SEM, n = 3). p < 0.05 (*), p < 0.001 (**), p < 0.0001 (***), compared with the DMSO-treated control by one-way ANOVA followed by Dunnett’s adjustment. DATS, Diallyl trisulfide, DEAB, diethylaminobenzaldehyde; BAAA, BODIPYTM-amino acetaldehyde; C means DMSO-treated control.

Article Snippet: Diallyl trisulfide Cancers 2024, 16, 378 3 of 19 (DATS, purity > 98%) was procured from LKT Laboratories (St. Paul, MN, USA), and RNase A was sourced from Promega (Madison, WI, USA).

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

Journal: Cancers

Article Title: Diallyl Trisulfide Induces ROS-Mediated Mitotic Arrest and Apoptosis and Inhibits HNSCC Tumor Growth and Cancer Stemness.

doi: 10.3390/cancers16020378

Figure Lengend Snippet: Figure 6. DATS administration inhibited the tumor growth of UMSCC-22B cells and cancer stem cells in vivo. (A) Body weights for control mice and those treated with DATS. (B) Representative tumor images of control mice and DATS-treated mice. (C) Average tumor weight in control and DATS-treated mice. In one mouse of the DATS group, the tumor regressed drastically on one side. (D) Average tumor volume as a function of time in control mice and DATS-treated mice (oral gavage administration, five times per week). There were four mice each in the control and DATS treatment group with tumor cells implanted on both the left and right flank of each mouse. (E) Representative images of Ki-67 immunohistochemical staining from control mouse tumor and DATS-treated mouse tumor (magnification ×200, scale bar = 100 µm). (F) Quantification of Ki-67 protein expression. The result shown is the mean H-score (n = 3 for control, and n = 3 for the DATS-treated group). (G) Representative flow histograms for ALDH1 activity from single cells isolated from control and DATS-treated mice tumors. The ALDH1 inhibitor DEAB was used as a control. (H) Quantitation of ALDH1 activity of respective groups. The results shown are mean ± SEM. The p-value was calculated by a two-sided Student’s t-test. DATS, Diallyl trisulfide, p < 0.05 (*), p < 0.0001 (***).

Article Snippet: Diallyl trisulfide Cancers 2024, 16, 378 3 of 19 (DATS, purity > 98%) was procured from LKT Laboratories (St. Paul, MN, USA), and RNase A was sourced from Promega (Madison, WI, USA).

Techniques: In Vivo, Control, Immunohistochemical staining, Staining, Expressing, Activity Assay, Isolation, Quantitation Assay

Journal: Cancers

Article Title: Diallyl Trisulfide Induces ROS-Mediated Mitotic Arrest and Apoptosis and Inhibits HNSCC Tumor Growth and Cancer Stemness.

doi: 10.3390/cancers16020378

Figure Lengend Snippet: Figure 7. Mechanism of DATS-induced cell cycle arrest, apoptosis, DNA damage, and inhibition of cancer stemness. DATS-induced ROS-mediated mitotic arrest, apoptosis, and DNA damage in HNSCC cells by altering the levels of the cell cycle regulatory proteins, apoptotic regulatory proteins, and DNA damage marker γ-H2AX. NAC (N-acetyl cysteine) pretreatment attenuates DATS-induced ROS-mediated mitotic arrest, apoptosis, and DNA damage. DATS treatment inhibited the cancer stem cell (CSC) fraction in vitro. DATS treatment inhibited the growth of HNSCC tumor xenograft and CSC fraction in vivo. Red arrow indicates decrease and green arrow indicates increase.

Article Snippet: Diallyl trisulfide Cancers 2024, 16, 378 3 of 19 (DATS, purity > 98%) was procured from LKT Laboratories (St. Paul, MN, USA), and RNase A was sourced from Promega (Madison, WI, USA).

Techniques: Inhibition, Marker, In Vitro, In Vivo