Journal: iScience

Article Title: Neuregulin-1/ErbB4 signaling modulates Plasmodium falciparum HRP2-induced damage to brain cortical organoids

doi: 10.1016/j.isci.2022.104407

Figure Lengend Snippet: HRP2 increases pro-inflammatory and Toll-like receptor (TLR) genes in cortical organoids. HRP2-induced inflammation-related genes were profiled Fold changes greater than 1.5 and p values < 0.05 calculated using Student’s t -test was considered to be a significant dysregulation. (A) A heatmap was generated showing up-and down-regulated genes. The effects of HRP2 on cortical organoids at day 50 in culture are shown by site (B), including cerebral cortex and glia, as well as by pathway (C), including TLR signaling. Further analysis revealed the relationship between upregulated genes and activation of TLR2 pathway (D). Western blot densitometry of organoid lysates (data represented as means ± SEM) confirms that HRP2 significantly induces TLR2 and TLR1 expression, whereas NRG1 reduces these effects (E, F). a: p < 0.05 compared to basal conditions; b: p < 0.05 compared to HRP2 treatment. One-way ANOVA and t -test were used. Please also see Figure S4 .

Article Snippet: Rabbit anti-human TLR1 , Cell Signaling , Cat #2209T RRID: AB_2303485.

Techniques: Generated, Activation Assay, Western Blot, Expressing

Journal: iScience

Article Title: Neuregulin-1/ErbB4 signaling modulates Plasmodium falciparum HRP2-induced damage to brain cortical organoids

doi: 10.1016/j.isci.2022.104407

Figure Lengend Snippet: HRP2 uses TLR1 and TLR2 to increase CXCL10 expression and cell death in iPSC Cell proliferation, apoptosis, and expression of inflammatory markers were assessed in iPSC treated with HRP2 after TLR1 and TLR2 blockage. (A) Proliferation assay showed that IPSC growth was partially restored, whereas their apoptosis (B) and necrosis (C) were reduced to the levels of untreated controls. TLR1 and TLR2 blocking in HRP2-treated iPSC has an antiapoptotic and anti-inflammatory effect, as it is confirmed by IHC: the expression of cleaved caspase 3 (D, F) and CXCL10 (E, G) is reduced. Data represented as means ± SEM a: p < 0.05 compared to basal conditions; b: p < 0.05 compared to HRP2 treatment. Statistical significance between groups was determined using t -test and analysis of variance.

Article Snippet: Rabbit anti-human TLR1 , Cell Signaling , Cat #2209T RRID: AB_2303485.

Techniques: Expressing, Proliferation Assay, Blocking Assay

Journal: iScience

Article Title: Neuregulin-1/ErbB4 signaling modulates Plasmodium falciparum HRP2-induced damage to brain cortical organoids

doi: 10.1016/j.isci.2022.104407

Figure Lengend Snippet:

Article Snippet: Rabbit anti-human TLR1 , Cell Signaling , Cat #2209T RRID: AB_2303485.

Techniques: Functional Assay, Recombinant, Plasmid Preparation, CCK-8 Assay, Bicinchoninic Acid Protein Assay, Isolation, Gene Expression, Western Blot, Software, Microscopy

Journal: Mediators of Inflammation

Article Title: Anti-Inflammatory Effects of IKK Inhibitor XII, Thymulin, and Fat-Soluble Antioxidants in LPS-Treated Mice

doi: 10.1155/2014/724838

Figure Lengend Snippet: The effects of IKK Inhibitor XII and antioxidant-rich diet on phosphorylation of RelA, IKK, and SAPK/JNK and on the expression of TLR4 and heat shock proteins in the splenic lymphocytes from inflammation-bearing mice. The animal's groups that was indicated in were used (1, control; 2, IB mice; 3, IB + Inh5; 4, IB + Inh10; 5, IB + Inh20; 6, IB + diet; 7, IB + diet + Inh20). Western blot analysis of extracts from isolated mice lymphocytes was provided using corresponding antibodies or anti-GAPDH antibody (bottom). Blot pictures show a single representative experiment from four independent experiments. Histograms below protein bands show protein levels calculated as mean relative units correspondingly to internal control and are the results of blots densitometry by program QAPA from four independent experiments. *Significantly different from control, P < 0.05. ∧ Significantly different from LPS-group, P < 0.05.

Article Snippet: After blocking, the membrane was exposed for 2 hr to antibodies against the following mouse proteins: HSP70 antibody (rabbit anti-mouse HSP 72, clone SPA-812, inducible form, StressGen), HSP90 antibody (rabbit anti-Hsp90 α [Hsp86], StressGen), phospho-NF- κ B antibody (phospho-NF- κ B p65 [Ser 536], #3031, Cell Signaling Technology, Danvers, MA), rabbit phospho- IKK α / β antibody II (Ser 176/180 (Cell Signaling Technology, USA), rabbit phospho-SAPK/JNK antibody to synthetic phospho-peptide SAPK/JNK, or rabbit TLR4 antibody (#2246, Cell Signaling Technology, USA).

Techniques: Phospho-proteomics, Expressing, Control, Western Blot, Isolation

Journal: Frontiers in Pharmacology

Article Title: Periplaneta americana extract alleviates steatohepatitis in a mouse model by modulating HMGB1-mediated inflammatory response

doi: 10.3389/fphar.2022.995523

Figure Lengend Snippet: GLC blocks M1 polarization by decreasing HMGB1 released by hepatocytes. (A) Immunofluorescence staining of HMGB I in mouse liver tissues. Scale bars, 50 μm. (B) Statistics of cytoplasmic HMGB1 positive cells of IF staining sections (C) Dataset (GEO database- GSE28619) of patients with alcoholic hepatitis were analyzed. Expression levels of M1 related markers (CD86, CXCL9, CXCL1 0 and TLR4) in HMGB1-High group and HMGB1-Low group. (D) Schematic flow chart of co-culture system. NCTC1469 cells induced by EtOH with or without treatment of GLC serum. Supemetant was collected for ELISA and culturing macrophages. The treatment of NCTC1469 cells was displayed in table (E) Supernatant HMGB1 levels of NCTC1469 cells induced by EtOH. ** P < 0.01 vs. the NCTC1469 cells with EtOH treatment. (F) Representative iNOS immunofluorescence staining and statistics of macrophages treated with condition medium (CM) of NCTC1469 cells. Scale bars, 50 μm. (G) Quantitative PCR analysis of J774A.1 cell cocultured with NCTCI469 cells. The data were obtained from three dependent experiments per group. * P < 0.05 vs. G#1, *** P < 0.001 vs. G#1.

Article Snippet: The membranes were washed, blocked and incubated with specific primary anti-rabbit antibodies against CYP2E1 (Proteintech, #19937-1-AP), GAPDH (Cell signaling Technology, #2118S), pNF-kB p65 (Wanleibio, #WL02169), NF-kB p65 (Wanleibio, #WL01273b), TLR4 (PTMBIO, #PTM-5192).

Techniques: Immunofluorescence, Staining, Expressing, Co-Culture Assay, Enzyme-linked Immunosorbent Assay, Real-time Polymerase Chain Reaction

Journal: Frontiers in Pharmacology

Article Title: Periplaneta americana extract alleviates steatohepatitis in a mouse model by modulating HMGB1-mediated inflammatory response

doi: 10.3389/fphar.2022.995523

Figure Lengend Snippet: GLC ameliorates progression of steatohepatitis via HMGB1-mediated TLR4/NF-KB pathway (A) Western blot analysis of indicated proteins expression. (B) The relative intensity of indicated proteins expression. The data were obtained from three dependent experiments per group. (C) Process of GLC inhibiting HMGB1-mediated macrophage activation by TLR4/NF-KB in the progression of steatohepatitis. Alcoholics and free fatty acids cause oxidative stress injury and lipid accumulation of hepatocytes. The dying/dead hepatocytes release DAMPs - HMGB1 to the extracellular. HMGB1 combined with TLR4 of macrophages to activate the inflammatory response through the NF-KB pathway, which aggravated the hepatocyte injury. GLC exerts hepatoprotective properties by reducing lipid accumulation and oxidative stress. It decreases the HMGB1 in extracellular and blocks TLR4/NF-KB mediated inflammatory cytokines expression of macrophages.

Article Snippet: The membranes were washed, blocked and incubated with specific primary anti-rabbit antibodies against CYP2E1 (Proteintech, #19937-1-AP), GAPDH (Cell signaling Technology, #2118S), pNF-kB p65 (Wanleibio, #WL02169), NF-kB p65 (Wanleibio, #WL01273b), TLR4 (PTMBIO, #PTM-5192).

Techniques: Western Blot, Expressing, Activation Assay