Journal: Cell Proliferation

Article Title: Tetrahedral Framework Nucleic Acid Relieves Sepsis‐Induced Intestinal Injury by Regulating M2 Macrophages

doi: 10.1111/cpr.13803

Figure Lengend Snippet: Treatment of tFNAs activated the Mertk signal pathway. (A) Representative images of immunofluorescent staining for tFNAs (red) and DAPI (blue) in the colon and ileum (Scale bar = 20 μm). (B) Immunohistochemical staining of Mertk in colon (scale bar: 100 μm). (C–F) Western blotting detection of Mertk, p‐Mertk, STAT1, p‐STAT1, SOCS1, SOCS3 expression in the colon. (G) Quantitative analysis of the protein expression levels. (H) Schematic diagram of the inhibition of Mertk altering macrophage activation by tFNAs. (I–L) Western blotting detection of Mertk, p‐Mertk, STAT1, p‐STAT1, SOCS1, SOCS3 expression in the colon. (M) Quantitative analysis of the protein expression levels. Data were represented as mean ± SD, * p < 0.05, ** p < 0.01.

Article Snippet: The antibody for ZO‐1 was from Wuhan Servicebio; the antibody for occludin was from Abcam; the antibodies for Mer, STAT1, p‐STAT1, SOCS1 and SOCS3 were from Cell Signalling Technology; the antibody for p‐Mer (Tyr749) was from Thermo Fisher Scientific; and UNC2250 was purchased from MCE.

Techniques: Staining, Immunohistochemical staining, Western Blot, Expressing, Inhibition, Activation Assay

Journal: Blood

Article Title: MDH1-mediated malate-aspartate NADH shuttle maintains the activity levels of fetal liver hematopoietic stem cells

doi: 10.1182/blood.2019003940

Figure Lengend Snippet: STAT3 transactivates Mdh1 expression to maintain FL-HSC activities. (A) Mdh1 luciferase reporter and different doses of Stat3 were cotransfected into 293T cells, followed by the determination of luciferase activities (n = 3). (B) ChIP assays were analyzed with 293T cells cotransfected with pGL4.27-mdh1 promoter vector and Stat3 plasmid or control plasmid. Input control and amplification of the Stat3-binding sequence of Mdh1 were determined by semiquantitative PCR. (C) Bisulfite sequencing analysis of the methylation patterns of the CpG islands in the Mdh1 promoter in CD45+ SoNar-high and -low FL hematopoietic cell fractions is shown. Each row represents a unique DNA clone; filled and open circles represent methylated and unmethylated CpGs, respectively. (D) Quantification data of methylated and unmethylated CpGs (n = 3). (E) Protein levels of pSTAT3, STAT3, and MDH1 were measured in CD45+ SoNar-high and -low FL cells. Ratios of pSTAT3/STAT3 and MDH1/actin were quantified and normalized against SoNar-high#1 cells. (F) Representative flow cytometric analysis of intracellular level of pSTAT3 of total CD45+ FL hematopoietic cells and FL-HSCs. Mean fluorescence intensity (MFI) of each group was shown (isotype control, yellow and green lines). (G) Quantification of MFI of intracellular level of pSTAT3 in panel F (n = 3). (H) Immunoblotting assay showed protein levels of pSTAT3, STAT3, and MDH1 in total CD45+ FL hematopoietic cells and FL-HSCs. Ratios of pSTAT3/STAT3 and MDH1/actin were quantified and normalized against total cells. (I) Representative flow cytometric analysis of intracellular level of pSTAT3 of CD45+ SoNar-high and -low FL-HSCs. MFI of each group is shown (isotype control, yellow and green lines). (J) Quantification of MFI of intracellular level of pSTAT3 in panel I (n = 3). (K) Protein level of MDH1 was measured in Stat3-overexpressed SoNar 32D cells and control cells by immunoblotting. Ratios of STAT3/actin and MDH1/actin were quantified and normalized against control cells. (L) Stat3-overexpressed SoNar 32D cells and control cells were evaluated for the ratios of SoNar fluorescence, and representative images are shown. Scale bar, 10 μm. (M) Quantification of the ratios of SoNar fluorescence in panel L. A total of 30 SoNar 32D cells were analyzed (n = 3). (N) Working model for the connections between FL-HSC activities and Mdh1-mediated malate-aspartate shuttle as indicated by SoNar. Data are represented as mean ± standard error of the mean. Student 2-tailed unpaired t test (G,J,M) and 2-way analysis of variance with Sidak’s multiple comparison test (D) were used for the comparison. *P < .05, ***P < .001.

Article Snippet: To test the effect of STAT3 inhibitor on MDH1 expression, purified Lin − Sca-1 + c-Kit + FL-HSCs and adult HSCs were treated with 10 or 20 μM of STAT3 inhibitor C188-9 (TargetMol) for 24 hours according to the previous study 37 and subjected to determination of pSTAT3, STAT3, or MDH1 level by western blot.

Techniques: Expressing, Luciferase, Plasmid Preparation, Control, Amplification, Binding Assay, Sequencing, Methylation Sequencing, Methylation, Fluorescence, Western Blot, Comparison

Journal: The Journal of allergy and clinical immunology

Article Title: The Immunology of AD and its Reversibility with Broad Spectrum and Targeted Therapies

doi: 10.1016/j.jaci.2017.01.011

Figure Lengend Snippet: Recent controlled trials in AD CRTH2 Prostaglandin D2 receptor 2; H4R Histamine H4 receptor; JAK Janus kinase; PDE4 Phosphodiesterase 4; TSLP thymic stromal lymphopoietin; TSLPR thymic stromal lymphopoietin receptor;

Article Snippet: Baricitinib , , JAK1/2 , Jak1/2 inhibitor – Oral small molecule , In Phase II , Eli Lilly , NCT02576938.

Techniques: