Journal: Cells

Article Title: NAD Modulates DNA Methylation and Cell Differentiation

doi: 10.3390/cells10112986

Figure Lengend Snippet: NAD induces myeloid differentiation in K562. ( a ) Wright Giemsa staining showing morphological changes between NAD-treated and control cells after four days. The figure is representative of five independent experiments ( b ) Representative image showing the increase in the surface markers CD15, CD14 and CD11b upon NAD treatment by flow cytometry. The table reports the mean % ±SD of CD14 + , CD15 + , CD11b + cells of five independent experiments. *: p < 0.05; **: p < 0.01; ***: p < 0.001 vs. vehicle ( n = 5) ( c ) NBT positive staining detected by small blue dots after counterstaining the cells with safranin. A magnification is shown in the rectangle. The pictures are representative of five independent experiments ( d ) Seahorse XF analysis of K562 mitochondrial stress response in cells treated with NAD or vehicle. The figure represents the mean of three biological replicates ( n = 3). Error bars indicate ± s.d.

Article Snippet: The cells were then stained with the Wright Giemsa solutions kit (CAMCO STAIN PAK Fort Lauderdale, FL, USA, pc#702) according to manufacturer’s instructions.

Techniques: Staining, Control, Flow Cytometry

Journal: Scientific Reports

Article Title: Si-based agent alleviated small bowel ischemia–reperfusion injury through antioxidant effects

doi: 10.1038/s41598-024-54542-7

Figure Lengend Snippet: Examination of goblet cells by mucin staining. ( a – d ) The representative microphotographs of the periodic acid-Schiff (PAS)-stained ( a , b ) or the Alcian blue-stained ( c , d ) or small intestine in the control (Con) ( a , c ) and Si groups ( b , d ). Scale bar: 40 µm ( a – d ). ( e , f ) The measured results of number of goblet cells in the damaged area in the Alcian blue-stained ( e ) or the PAS-stained ( f ) small intestine. Bar chart indicates the mean values of Goblet cells per villi. White: Con group. Grey: Si group. Data are expressed as mean. Standard error of the mean (SEM) of three (Con) or five mice (Si) per group. * p < 0.05 versus Con group, determined by Student’s paired t -test.

Article Snippet: Samples were stained using a PAS staining solution kit (ScyTek Laboratories, Inc., Logan, UT, USA) according to the manufacturer’s instructions.

Techniques: Staining, Control

Journal: bioRxiv

Article Title: NAD modulates DNA methylation and cell differentiation

doi: 10.1101/2020.07.31.231555

Figure Lengend Snippet: ( A ) Wright Giemsa staining showing morphological changes between NAD-treated and control cells after four days. ( B ) Increase in the surface markers CD15, CD14 and CD11b upon NAD treatment ( C ) NBT positive staining detected by small blue dots after counterstaining the cells with safranin. A magnification is shown in the rectangle. ( D ) Seahorse XF analysis of K562 mitochondrial stress response in cells treated with NAD or vehicle. The figure represents the mean of two biological replicates (n=2). Error bars indicate ± S.D. ( E ) Model showing the molecular mechanism of CEBPA gene reactivation by NAD. CEBPA is epigenetically silenced in K562. DNMT1 ensure a constant methylated status of CEBPA promoter (upper part). The NAD supplementation to K562 cell culture, boosts PARP1 to produce ADP-ribose polymers leading to DNMT1 inhibition (bottom part). The ultimate effect is CEPBA re-activated transcription.

Article Snippet: The cells were then stained with the Wright Giemsa solutions kit (CAMCO STAIN PAK, pc#702) according to manufacturer’s instructions.

Techniques: Staining, Methylation, Cell Culture, Inhibition

Journal: Med (New York, N.y.)

Article Title: Intestinal barrier dysfunction plays an integral role in arthritis pathology and can be targeted to ameliorate disease

doi: 10.1016/j.medj.2021.04.013

Figure Lengend Snippet: Mice with arthritis display intestinal permeability and intestinal inflammation (A) Concentration of LBP in serum of K/BxN mice at marked ages and naive C57BL/6 control mice (n = 3/group), as measured by ELISA. (B) Correlation of serum LBP concentration with increase in joint size in 4- to 5-week-old K/BxN mice (n = 8). (C) Concentration of FITC-dextran in serum of naive C57BL/6 control mice (CTRL) (n = 9) and K/BxN mice with early arthritis (n = 7) and with established arthritis (n = 9). (D–J) Representative (D) ZO-1 staining, (E) mean ZO-1 intensity, (F) H&E staining, (G) histological scores, (H) MUC2 staining, (I) mean MUC2 intensity, and (J) goblet cell counts from periodic acid-Schiff (PAS) staining for SI of K/BxN mice throughout arthritis development and compared to naive C57BL/6 control mice. ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001; ∗∗∗∗p < 0.0001. (A, C, E, I, and J) One-way ANOVA, (B) Pearson’s correlations, and (G) two-way ANOVA. For all panels, one out of two experiments is shown. Data represent mean ± S.E.M.

Article Snippet: The cryo-sections (6 μm) were fixed with 10% neutral buffered formalin solution (Sigma) for 10 min (RT) and stained for mucin and goblet cell counts using a PAS staining kit according to the manufacturer’s instructions (ab150680, abcam).

Techniques: Permeability, Concentration Assay, Enzyme-linked Immunosorbent Assay, Staining

Journal: Med (New York, N.y.)

Article Title: Intestinal barrier dysfunction plays an integral role in arthritis pathology and can be targeted to ameliorate disease

doi: 10.1016/j.medj.2021.04.013

Figure Lengend Snippet:

Article Snippet: The cryo-sections (6 μm) were fixed with 10% neutral buffered formalin solution (Sigma) for 10 min (RT) and stained for mucin and goblet cell counts using a PAS staining kit according to the manufacturer’s instructions (ab150680, abcam).

Techniques: Recombinant, Methylation, Isolation, SYBR Green Assay, Purification, Enzyme-linked Immunosorbent Assay, Binding Assay, Staining, Software, Lysis, Plasmid Preparation