Journal: International Journal of Molecular Sciences

Article Title: Melatonin Exerts Prominent, Differential Epidermal and Dermal Anti-Aging Properties in Aged Human Eyelid Skin Ex Vivo

doi: 10.3390/ijms242115963

Figure Lengend Snippet: Overview of the impact of melatonin administration on key skin aging biomarkers. ↑ = increased, ↔ = no change, ↓ = decreased, compared to the vehicle. Mann–Whitney or Student’s t -test, * p < 0.5, ** p < 0.01.

Article Snippet: , γH2A.x , 4% PFA, 10 min at RT , 10% goat serum in PBS , rabbit monoclonal anti-phospho-histone H2A.X (Ser139) (γH2A.X) (1:1500; Cell Signaling, #2577S) , goat monoclonal anti-rabbit IgG-Alexa Fluor 555 (1:400; Invitrogen, A21428).

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: Melatonin Exerts Prominent, Differential Epidermal and Dermal Anti-Aging Properties in Aged Human Eyelid Skin Ex Vivo

doi: 10.3390/ijms242115963

Figure Lengend Snippet: List of antibodies and immunohistochemical staining treatments used in the study.

Article Snippet: , γH2A.x , 4% PFA, 10 min at RT , 10% goat serum in PBS , rabbit monoclonal anti-phospho-histone H2A.X (Ser139) (γH2A.X) (1:1500; Cell Signaling, #2577S) , goat monoclonal anti-rabbit IgG-Alexa Fluor 555 (1:400; Invitrogen, A21428).

Techniques: Immunohistochemistry, Staining, Blocking Assay, Amplification, Avidin-Biotin Assay

Journal: iScience

Article Title: WIP1 is a novel specific target for growth hormone action

doi: 10.1016/j.isci.2023.108117

Figure Lengend Snippet:

Article Snippet: Rabbit anti-Phospho-Histone H2A.X (Ser139) , Cell Signaling Technology , Cat# 9718; RRID: AB_10121789.

Techniques: Virus, shRNA, Recombinant, Protease Inhibitor, SYBR Green Assay, Enzyme-linked Immunosorbent Assay, DC Protein Assay, Extraction, Immunoprecipitation, Single Cell Gel Electrophoresis, Derivative Assay, Generated, Software, Imaging, Real-time Polymerase Chain Reaction, Microscopy

Journal: Cell reports

Article Title: Vitamin K-dependent carboxylation regulates Ca 2+ flux and adaptation to metabolic stress in β cells

doi: 10.1016/j.celrep.2023.112500

Figure Lengend Snippet: (A and B) INS-1 832/3 cells were cultured with VK 1 (22 μM) or vehicle for 48 h before being cultured for 24 h in medium containing 25 mM glucose and thapsigargin. (A) Cellular fitness was analyzed by western blot using cleaved caspase-3 and p(Ser139)-histone H2A.X antibodies. Quantification was performed using arbitrary densitometry units of cleaved caspase-3 and p(Ser139)-histone H2A.X signals obtained in samples treated with 20 nM thapsigargin over b-actin signals. (B) GGCX γ-carboxylation assessed by anti-Gla IP followed by Western blot using anti-GGCX antibodies. Quantification was performed using arbitrary densitometry units of GGCX and Gla-GGCX signals over b-actin signals. (C and D) Islets from C57BL/6J mice were cultured in medium containing either 5 or 15 mM glucose, and expression was analyzed by (C) qPCR (n = 3–4) and (D) western blot. (E) INS-1 832/3 cells were cultured in medium containing 2.5 or 25 mM glucose in the presence of VK 1 (22 μM) or vehicle, and GGCX γ-carboxylation was assessed by anti-Gla IP followed by western blot. (F and G) Two-months old Wistar rats were infused during 3 days with saline or glucose. (F) Average blood glucose level for each mouse. (G) Gene expression analyzed by qPCR (n = 3–4). (H and I) Human islets from non-diabetic cadaveric donors were cultured in the presence of VK 1 (22 μM) in medium containing either 5 or 15 mM glucose for 3 days. (H) Representative western blot experiment with islets from donor R288. (I) γ-Carboxylation was quantified using arbitrary densitometry units of Gla signals over b-actin signals. Data from 15 mM glucose-treated samples were normalized over 5 mM glucose treatment (n = 3). Results represent the mean ± SEM. Unpaired, 2-tailed Student’s t test was used in (C), (F), (G), and (I). ***p < 0.001, **p < 0.01, *p < 0.05. See also and .

Article Snippet: rabbit anti-phospho(Ser139)-Histone H2A.X , Cell Signaling Technology , Cat# 9718; RRID:AB_2118009.

Techniques: Cell Culture, Western Blot, Expressing

Journal: Cell reports

Article Title: Vitamin K-dependent carboxylation regulates Ca 2+ flux and adaptation to metabolic stress in β cells

doi: 10.1016/j.celrep.2023.112500

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: rabbit anti-phospho(Ser139)-Histone H2A.X , Cell Signaling Technology , Cat# 9718; RRID:AB_2118009.

Techniques: Recombinant, SYBR Green Assay, TUNEL Assay, Enzyme-linked Immunosorbent Assay, Plasmid Preparation, RNA Sequencing Assay, Expressing, Clone Assay, Sequencing, Knock-Out, Software, CRISPR, Transfection, Purification

Journal: International Journal of Molecular Sciences

Article Title: Less Severe Polymicrobial Sepsis in Conditional mgmt -Deleted Mice Using LysM-Cre System, Impacts of DNA Methylation and MGMT Inhibitor in Sepsis

doi: 10.3390/ijms241210175

Figure Lengend Snippet: The characteristics of bone marrow-derived macrophages (BMDM) from mgmt control (mgmt fl/fl ; LysM-Cre -/- ) or mgmt null (mgmt fl/fl ; LysM-Cre cre/- ) mice at 24 h after activation by lipopolysaccharide (LPS) as indicated by supernatant cell-free DNA ( A ), malondialdehyde (MDA; a reactive stress molecule) ( B ), and immunofluorescent stained for DNA break (phospho-histone H2A.X; green color) and actin filament (red color) in intensity score and representative pictures ( C , D ) are demonstrated. Triplicated independent experiments were performed. Mean ± SEM with the one-way ANOVA followed by Tukey’s analysis was used. #, p ˂ 0.05 vs mgmt control DMEM; *, p ˂ 0.05 between the indicated groups.

Article Snippet: Fixed samples were blocked with 2% bovine serum albumin in 1X TBS for 1 h at room temperature and then incubated overnight at 4 °C with phospho-histone H2A.X (Ser139) (20E3) rabbit mAb (Cell signaling).

Techniques: Derivative Assay, Activation Assay, Staining