Journal: eLife

Article Title: Functional identification of soluble uric acid as an endogenous inhibitor of CD38

doi: 10.7554/eLife.96962

Figure Lengend Snippet:

Article Snippet: Commercial assay or kit , Wright-Giemsa Stain Kit , ScyTek Laboratories , Cat#WGK-1 , .

Techniques: Recombinant, Enzyme-linked Immunosorbent Assay, Staining, Colorimetric Assay, Software, Activity Assay

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: International Journal of Molecular Medicine

Article Title: HMGB1 participates in LPS-induced acute lung injury by activating the AIM2 inflammasome in macrophages and inducing polarization of M1 macrophages via TLR2, TLR4, and RAGE/NF-κB signaling pathways

doi: 10.3892/ijmm.2019.4402

Figure Lengend Snippet: Extracellular HMGB1 affects the inflammatory response in lung tissues in ALI. Histopathological changes and injury scores in the left lung are shown for different groups: (A) Control group, LPS group, Anti-HMGB1 group and LPS + anti-HMGB1 group (H&E staining, ×200 magnification). (B) Control group, LPS group, rHMGB1 group and LPS + rHMGB1 group (H&E staining, ×200 magnification). Effects of (C) anti-HMGB1 and (D) rHMGB1 on the lung W/D ratio of LPS-induced ALI mice. Effects of (E) anti-HMGB1 and (F) rHMGB1 on MPO activity in lung tissues and infiltration of inflammatory cells in BALF of LPS-induced ALI mice. All the experiments were repeated more than three times (n=4-6 mice per each group). Data presented is from a selected representative experiment. All data are expressed as the mean ± standard deviation. * P<0.05, ** P<0.01 and *** P<0.001 vs. LPS group; # P<0.05 vs. control group. Extracellular HMGB1 affects the inflammatory response in lung tissues in ALI. Effects of (G) anti-HMGB1 and (H) rHMGB1 on MPO activity in lung tissues and infiltration of inflammatory cells in BALF of LPS-induced ALI mice. Effects of (I) anti-HMGB1 and (J) rHMGB1 on the ELISA results of secretion of inflammatory cytokines (TNF-α, IL-6, MCP-1, IL-1β, and IL-18) in BALF. All the experiments were repeated more than three times (n=4-6 mice per each group). Data presented is from a selected representative experiment. All data are expressed as the mean ± standard deviation. * P<0.05, ** P<0.01 and *** P<0.001 vs. LPS group; # P<0.05 vs. control group. LPS, lipopolysaccharide; IL, interleukin; BALF, bronchoalveolar lavage fluid; MCP, myeloper-oxidase; TNF, tumor necrosis factor; ALI, acute lung injury; rHMGB1, recombinant High mobility group box 1; H&E, hematoxylin and eosin; W/D, wet to dry.

Article Snippet: Total and differential inflammatory cells in BALF were counted using Wright-Giemsa staining (3-5 drops of methanol fixing agent was added to the cells at room temperature for 1 min; Wright-Giemsa stain was then added at room temperature for 8 min, and the cells were finally flushed with running water for 30-60 sec; Wright-Giemsa Stain kit; WGK-1; ScyTek; Shenzhen Xinbosheng Bioengineering Institute) for cytospin preparations.

Techniques: Control, Staining, Activity Assay, Standard Deviation, Enzyme-linked Immunosorbent Assay, Recombinant

Journal: International Journal of Molecular Medicine

Article Title: HMGB1 participates in LPS-induced acute lung injury by activating the AIM2 inflammasome in macrophages and inducing polarization of M1 macrophages via TLR2, TLR4, and RAGE/NF-κB signaling pathways

doi: 10.3892/ijmm.2019.4402

Figure Lengend Snippet: Reduced inflammatory response in lung tissue in ALI by administration of TLR2/4 and RAGE antagonists. Histopathological changes and injury scores in left lung are illustrated for different groups: (A) Control group, LPS group, LPS-RS group, and LPS + LPS-RS group (H&E staining, ×200). (B) Control group, LPS group, FPS-ZM1 group and D: LPS+FPS-ZM1 group (H&E staining, ×200). Effects of (C) LPS-RS and (D) FPS-ZM1 on the lung W/D ratio of LPS-induced ALI mice. Effects of LPS-RS and on (E) MPO activity in lung tissues and (F) infiltration of inflammatory cells in BALF of LPS-induced ALI mice. All experiments were repeated more than three times (n=4-6 mice per each group). Data presented is from a representative experiment. All data are expressed as the mean ± standard deviation. * P<0.05, ** P<0.01 and *** P<0.001 vs. LPS group. Reduced inflammatory response in lung tissue in ALI by administration of TLR2/4 and RAGE antagonists. Effects of FPS-ZM1 on (G) MPO activity in lung tissues and (H) infiltration of inflammatory cells in BALF of LPS-induced ALI mice. ELISA results of secretion of inflammatory cytokines (TNF-α, IL-6, MCP-1, IL-1β and IL-18) treated with (I) LPS-RS and (J) FPS-ZM1 in BALF. All experiments were repeated more than three times (n=4-6 mice per each group). Data presented is from a representative experiment. All data are expressed as the mean ± standard deviation. * P<0.05, ** P<0.01 and *** P<0.001 vs. LPS group. LPS, lipopolysaccharide; IL, interleukin; MCP, myeloperoxidase; TNF, tumor necrosis factor; ALI, acute lung injury; rHMGB1, recombinant High mobility group box 1; RT-q, reverse transcription-quantitative; NF, nuclear factor; BMMs, bone-marrow derived macrophages; TLR, toll-like receptor; RAGE, receptor for advanced glycation end products; BALF, bronchoalveolar lavage fluid; H&E, hematoxylin and eosin; RS, Rhodobacter sphaeroides ; Mac, macrophage; Neu, neutrophil; Lym, lymphocyte.

Article Snippet: Total and differential inflammatory cells in BALF were counted using Wright-Giemsa staining (3-5 drops of methanol fixing agent was added to the cells at room temperature for 1 min; Wright-Giemsa stain was then added at room temperature for 8 min, and the cells were finally flushed with running water for 30-60 sec; Wright-Giemsa Stain kit; WGK-1; ScyTek; Shenzhen Xinbosheng Bioengineering Institute) for cytospin preparations.

Techniques: Control, Staining, Activity Assay, Standard Deviation, Enzyme-linked Immunosorbent Assay, Recombinant, Reverse Transcription, Derivative Assay