Journal: Advanced Science

Article Title: VDLIN: A Deep Learning‐Based Platform for Methylcobalamin‐Inspired Immunomodulatory Compound Screening

doi: 10.1002/advs.202413775

Figure Lengend Snippet: Co7 induces Ifnb1 expression via the TLR4 signaling pathway. A) Volcano plot illustrating the distribution of differentially expressed genes (DEGs) between the Co7 and DMSO treatment groups after 3 h in RAW 264.7 cells. Fold changes are presented as log 2 transformations. Red dots represent DEGs upregulated in the Co7 group. B) Protein‐protein interaction (PPI) network analysis of differentially expressed genes (DEGs) in the Co7‐treated group compared to the DMSO group, revealing significant enrichment in pathways associated with the innate immune response and type I interferon signaling. C) KEGG pathway enrichment analysis of DEGs induced by Co7, highlighting associations with innate immune response pathways. D) Co7 (50 µmol/L) exhibiting strong antiviral effects against VSV in RAW 264.7 and HT29 cells. E) Co7 significantly reduced the inflammatory response induced by LPS, VSV, EMCV, and HSV in RAW 264.7 cells. F) Volcano plot representing the differential gene expression analysis between Co7‐ and LPS‐treated RAW 264.7 cells after 3 h of treatment. Fold changes are displayed as log 2 transformations. Red dots indicate genes upregulated in the Co7 group, while blue dots represent downregulated genes compared to LPS treatment. G) Western blot analysis demonstrating that Co7 inhibited the expression of iNOS and COX2, as well as the phosphorylation of NF‐κB‐P65 at the protein level in RAW 264.7 cells. H) Co7 significantly reduced the mortality rate in mice (n = 10 per group) following LPS challenge (20 mg/kg), compared to the PBS control group. RT‐qPCR data were presented as means ± SEM from three independent experiments. Statistical significance was determined using one‐way ANOVA with Bonferroni's multiple comparisons test (left three panels in E), paired‐samples t‐test (D, right panel of EMCV and HSV in E), or the log‐rank test (H). * P < 0.05, ** P < 0.01, and *** P < 0.001.

Article Snippet: Each inhibitor was dissolved in anhydrous DMSO and diluted to its respective working concentration: C29 (10 μM, S6597, Selleck) served as a TLR2 inhibitor; Procyanidin B1 (30 μM, HY‐N0795, MedChemExpress) acted as a TLR4 inhibitor; MyD88‐IN‐1 (30 μM, HY‐149992, MedChemExpress) was used to inhibit MyD88; Pepinh‐TRIF TFA (30 μM, HY‐P2565, MedChemExpress) functioned as a TRIF inhibitor; and GSK8612 (5 μM, T5540, TargetMol) inhibited TBK1/IKKε.

Techniques: Expressing, Gene Expression, Western Blot, Phospho-proteomics, Control, Quantitative RT-PCR

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: FIGURE 4 | 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: Viruses

Article Title: ZIKV Strains Elicit Different Inflammatory and Anti-Viral Responses in Microglia Cells

doi: 10.3390/v15061250

Figure Lengend Snippet: Infection of BV2 microglia cells with ZIKV PE243 and ZIKV MR766 strains differentially modulate the pro-inflammatory miRNA-155 and anti-inflammatory and anti-viral factors. BV2 cells were infected with both ZIKV PE243 or ZIKV MR766 , with an MOI of 1, at 12, 24, 48, and 72 hpi using mock-treated (Mock) as a negative control and LPS + ATP (LPS) as a positive control. ( a ) miRNA-155, ( b ) SOCS1, ( d ) SOCS3 and ( e ) SHIP1 mRNA expression levels were measured by RT-qPCR. ∆∆Ct plotted results were normalized to hprt1 and mock values for mRNAs, and U6 and mock for miRNAs. ( c ) The protein SOCS1 expressions were analyzed by Western analysis. The membrane was stained with anti-SOCS1 and anti-β-actin as a loading control. Statistical analysis was performed by One-Way ANOVA, considering * p < 0.05, ** p < 0.01, and *** p < 0.001 as significant differences.

Article Snippet: Primary antibodies against PPAR-γ (Thermo Fisher Scientific Inc., PA3-821A) and SOCS1 (Boster Biological Technology Co. Ltd, Pleasanton, CA, USA, PA1074) were added to the membranes, and then secondary antibodies against mouse (Cell Signaling Technology, 7076S, Danvers, MA, USA) and rabbit (KPL, 04-15-06) were used.

Techniques: Infection, Negative Control, Positive Control, Expressing, Quantitative RT-PCR, Western Blot, Membrane, Staining, Control

Journal: Viruses

Article Title: ZIKV Strains Elicit Different Inflammatory and Anti-Viral Responses in Microglia Cells

doi: 10.3390/v15061250

Figure Lengend Snippet: Comparison of inflammatory, anti-inflammatory, and anti-viral marker expressions by BV2 microglial cell line after infection with Brazilian and African ZIKV strains (ZIKV PE243 and ZIKV MR766 ).

Article Snippet: Primary antibodies against PPAR-γ (Thermo Fisher Scientific Inc., PA3-821A) and SOCS1 (Boster Biological Technology Co. Ltd, Pleasanton, CA, USA, PA1074) were added to the membranes, and then secondary antibodies against mouse (Cell Signaling Technology, 7076S, Danvers, MA, USA) and rabbit (KPL, 04-15-06) were used.

Techniques: Comparison, Marker, Infection

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: