Journal: Current Research in Immunology

Article Title: Gamma-tocotrienol modifies methylation of HOXA10, IRF4 and RORα genes in CD4 + T-lymphocytes: Evidence from a syngeneic mouse model of breast cancer ☆

doi: 10.1016/j.crimmu.2021.10.001

Figure Lengend Snippet: Comparing percentage of DNA methylation in CD4 + T-lymphocytes from ( A ) control (no tumour) or ( B ) tumour-induced mice fed with (A) vehicle (soy oil) or (B) γT3 (γT3 in soy oil). Genomic DNA was extracted from CD4 + T-lymphocytes (QIAGEN DNeasy Blood and Tissue mini kit) isolated from peripheral blood at autopsy. The DNA was analysed for modification in the DNA methylation of the 22 genes that were annotated in the commercial DNA methylation array (EpiTech Methyl II signature PCR for murine T-helper differentiation array plate, SABioscience, USA). Data is represented as mean percentage of methylation ± standard deviation (SD) and is representative of at least three independent mice. (*P < 0.05 versus vehicle, tumour).

Article Snippet: Gamma-tocotrienol (γT3) was provided by Davos Life Sciences, Pte Ltd, Singapore.

Techniques: DNA Methylation Assay, Control, Isolation, Modification, Methylation, Standard Deviation

Journal: Current Research in Immunology

Article Title: Gamma-tocotrienol modifies methylation of HOXA10, IRF4 and RORα genes in CD4 + T-lymphocytes: Evidence from a syngeneic mouse model of breast cancer ☆

doi: 10.1016/j.crimmu.2021.10.001

Figure Lengend Snippet: Comparing percentage of DNA methylation in CD4 + T-lymphocytes from tumour-induced mice fed with (A) vehicle (soy oil) or (B) γT3 (γT3 in soy oil). Genomic DNA was extracted from CD4 + T-lymphocytes (QIAGEN DNeasy Blood and Tissue mini kit) isolated from peripheral blood at autopsy. The DNA was analysed for modification in the DNA methylation of the 22 genes that were annotated in the commercial DNA methylation array (EpiTech Methyl II signature PCR for murine T-helper differentiation array plate, SABioscience, USA). Data is represented as mean percentage of methylation ± standard deviation (SD) and is representative of at least three independent mice. (*P < 0.05 versus vehicle, tumour).

Article Snippet: Gamma-tocotrienol (γT3) was provided by Davos Life Sciences, Pte Ltd, Singapore.

Techniques: DNA Methylation Assay, Isolation, Modification, Methylation, Standard Deviation

Journal: Cell Proliferation

Article Title: Anti‐proliferative effects of γ‐tocotrienol on mammary tumour cells are associated with suppression of cell cycle progression

doi: 10.1111/j.1365-2184.2009.00657.x

Figure Lengend Snippet: Effects of γ‐tocotrienol on cyclin D1 and CDK levels. +SA cells were plated at a density of 1 × 106 cells/100 mm culture dishes in serum‐free defined control media, to allow cells to attach. The following day they were divided into control and treatment groups, media were removed, cells were washed and then fed fresh mitogen‐free media containing 0 or 4 μmγ‐tocotrienol (γT3), to establish cell cycle synchronization in all groups. After 48 h incubation period, media were removed and cells were fed their respective control or treatment media containing 100 ng/ml EGF. Cells were then collected at 0, 2, 4, 6, 12 and 24 h after mitogen stimulation and whole cell lysates were prepared for Western blot analysis of intracellular levels of cyclin D1, CDK2, CDK4, CDK6 and actin (α, β, γ) proteins. For each experiment, samples were electrophoresed through SDS–polyacrylamide minigels run simultaneously and proteins from two minigels were transblotted on to a single PVDF membrane, followed by Western blot analysis. Each Western blot image is a representative example of data obtained from experiments that were repeated at least three times. Scanning densitometric analysis was performed for each blot and net intensity of each band was normalized with corresponding actin values, as shown in bar graphs adjacent to their respective Western blot images. Vertical bars in the graph indicate relative protein expression in control and treatment groups over 24 h after mitogen (EGF) stimulation.

Article Snippet: Purified γ‐tocotrienol (>98% purity) was generously provided by Carotech Bhd. (Ipoh, Malaysia).

Techniques: Control, Incubation, Western Blot, Membrane, Expressing

Journal: Cell Proliferation

Article Title: Anti‐proliferative effects of γ‐tocotrienol on mammary tumour cells are associated with suppression of cell cycle progression

doi: 10.1111/j.1365-2184.2009.00657.x

Figure Lengend Snippet: Effects of γ‐tocotrienol on CKI levels. +SA cells were plated at a density of 1 × 106 cells/100 mm culture dishes and fed serum‐free defined control media to allow cells to attach. The following day, cells were divided into control and treatment groups, media were removed, cells were washed and then fed fresh mitogen‐free media containing 0 or 4 μmγ‐tocotrienol (γT3), to establish cell cycle synchronization between all groups. After 48 h incubation, media were removed and cells in all groups were fed their respective control or treatment media containing 100 ng/ml EGF. Cells were collected at various time points over the next 24 h period and whole cell lysates were prepared for Western blot analysis of p21, p27, p15 and actin (α, β, γ) proteins. For each experiment, samples were electrophoresed through SDS–polyacrylamide minigels run simultaneously and proteins from two minigels were transblotted on to a single PVDF membrane, followed by Western blot analysis. Each Western blot image is a representative example of data obtained from experiments that were repeated at least three times. Scanning densitometric analysis was performed for each blot and net intensity of each band was normalized to corresponding actin values, as shown in bar graphs adjacent to their respective Western blot images. Vertical bars in the graph indicate relative protein expression in control and treatment groups over 24 h after mitogen (EGF) stimulation.

Article Snippet: Purified γ‐tocotrienol (>98% purity) was generously provided by Carotech Bhd. (Ipoh, Malaysia).

Techniques: Control, Incubation, Western Blot, Membrane, Expressing

Journal: Cell Proliferation

Article Title: Anti‐proliferative effects of γ‐tocotrienol on mammary tumour cells are associated with suppression of cell cycle progression

doi: 10.1111/j.1365-2184.2009.00657.x

Figure Lengend Snippet: Effects of γ‐tocotrienol on retinoblastoma (Rb) phosphorylation. +SA cells were plated at a density of 1 × 106 cells/100 mm culture dishes and fed serum‐free defined control media to allow cells to attach. The following day, cells were divided into control and treatment groups, media were removed, cells washed, and then fed fresh mitogen‐free media containing 0 or 4 μmγ‐tocotrienol (γT3), to establish cell cycle synchronization in all groups. After 48 h incubation, media were removed and cells in all groups were fed their respective control or treatment media containing 100 ng/ml EGF. Cells were collected at specific times over the next 24 h period and whole cell lysates were prepared for Western blot analysis of intracellular levels of phospho‐Rb (ser780), phospho‐Rb (ser807/811) and actin (α, β, γ) proteins. For each experiment, samples were electrophoresed through SDS–polyacrylamide minigels run simultaneously and proteins from two minigels were transblotted on to a single PVDF membrane, followed by Western blot analysis. Each Western blot image is a representative example of data obtained from experiments that were repeated at least three times. Scanning densitometric analysis was performed for each blot and net intensity of each band was normalized with corresponding actin values, as shown in bar graphs adjacent to their respective Western blot images. Vertical bars in the graph indicate relative protein expression in control and treatment groups over 24 h after mitogen (EGF) stimulation.

Article Snippet: Purified γ‐tocotrienol (>98% purity) was generously provided by Carotech Bhd. (Ipoh, Malaysia).

Techniques: Phospho-proteomics, Control, Incubation, Western Blot, Membrane, Expressing

Journal: Cell Proliferation

Article Title: Anti‐proliferative effects of γ‐tocotrienol on mammary tumour cells are associated with suppression of cell cycle progression

doi: 10.1111/j.1365-2184.2009.00657.x

Figure Lengend Snippet:   Effects of 0–4 μmγ‐tocotrienol on the highly malignant +SA mammary epithelial cell proliferation over a 96 h treatment period. Cells in all treatment groups were initially plated (day 0) at a density of 5 × 104 cells/well in 24‐well culture plates. Vertical bars indicate mean viable cell number/well ± SEM for six replicates in each treatment group. *P < 0.05, as compared to vehicle‐treated cells in the control group.

Article Snippet: Purified γ‐tocotrienol (>98% purity) was generously provided by Carotech Bhd. (Ipoh, Malaysia).

Techniques: Control