Journal: iScience

Article Title: β-chitosan attenuates hepatic macrophage-driven inflammation and reverses aging-related cognitive impairment.

doi: 10.1016/j.isci.2024.110766

Figure Lengend Snippet: Figure 3. b-chitosan supplementation alleviated neuroinflammation by suppressing the microglia activation in the hippocampus of aged mouse (A) Western blot analyses of VCAM1, CD68, IL-1b, IBA1 and TNF-a expressions in the hippocampi of young (4 months old) and aged (13 months old) mice after administration. (B) Quantification analysis for the protein expressions in (A). (C) Confocal images of VCAM1 (red) and Lectin (green) staining in the CA1 of mice treated with b-chitosan. DAPI labels cell nucleus. Arrowheads indicate VCAM1+Lectin+ vessels. Scale bars, 50 mm. (D) Representative confocal images show the co-localization of IBA1 (red), CD68 (green) and DAPI (blue) in the CA1 of mice. Arrowheads indicate CD68+IBA1+ cells. Scale bars, 50 mm.

Article Snippet: Themembranes were blocked with 5% no-fat milk/TBST at room temperature for 1 h and then incubated with the primary antibody (1:1000 dissolved in TBST, see key resources table) at 4 C for at least 17 h. The secondary antibodies (1:5000, ThermoFisher Scientific, 31234, Rabbit; 31166, Mouse) of the corresponding species were incubated at room temperature for 1 h after the first antibody was recovered, visualized by using ECL Western Boltting Substrate. iScience 27, 110766, September 20, 2024 19 The primary antibodies for western blot, mice: VCAM1 (Abcam, ab134047, RRID:AB_2721053), CD68 (ProteinTech, 28058-1-AP), IL-1b (Santa Cruz Biotechnology, SC-52012), TNF-a (Affinity, AF7014), IBA1 (ProteinTech, 10904-1-AP), NLRP3 (Affinity, DF7438), MMP9 (Abmart, TA0220), IL-6 (Abmart, TD6087); cell: iNOS (Affinity, AF0199), CD86(Affinity, DF6332), TLR4 (ProteinTech, 19811-1-AP); JNK (ProteinTech, 24164-1-AP); p38 (ProteinTech, 14064-1-AP); p65 (ProteinTech, 10745-1-AP); p-JNK (Affinity, AF3318); p-p38 (Affinity, AF4001); p-p65 (Affinity, AF2006), TNF-a (Affinity, AF7014), IL-1b (Affinity, AF5103); zebrafish: IL-1b (ProteinTech, 16806-1-AP). b-tubulin (Affinity, AF7018), GAPDH (Affinity, AF7021), b-actin (Affinity, AF7011).

Techniques: Activation Assay, Western Blot, Staining

Journal: iScience

Article Title: β-chitosan attenuates hepatic macrophage-driven inflammation and reverses aging-related cognitive impairment.

doi: 10.1016/j.isci.2024.110766

Figure Lengend Snippet: Figure 5. b-chitosan supplementation prevented LPS-induced neuroinflammation (A) Representative confocal images show the co-localization of VCAM1+Lectin+ vessels (up) and CD68+IBA1+ cells (down) in the CA1 of LPS-challenged young mice (4 months old) after administration. DAPI labels cell nucleus. Arrowheads indicate VCAM1+Lectin+ vessels or CD68+IBA1+ cells, respectively. Scale bars, 50 mm. (B) Immunofluorescence analysis of the co-localization of VCAM1+Lectin+ vessels and CD68+IBA1+ cells in the DG of LPS-injected young mice. DAPI labels cell nucleus. Arrowheads indicate the positive signal. Scale bars, 50 mm.

Article Snippet: Themembranes were blocked with 5% no-fat milk/TBST at room temperature for 1 h and then incubated with the primary antibody (1:1000 dissolved in TBST, see key resources table) at 4 C for at least 17 h. The secondary antibodies (1:5000, ThermoFisher Scientific, 31234, Rabbit; 31166, Mouse) of the corresponding species were incubated at room temperature for 1 h after the first antibody was recovered, visualized by using ECL Western Boltting Substrate. iScience 27, 110766, September 20, 2024 19 The primary antibodies for western blot, mice: VCAM1 (Abcam, ab134047, RRID:AB_2721053), CD68 (ProteinTech, 28058-1-AP), IL-1b (Santa Cruz Biotechnology, SC-52012), TNF-a (Affinity, AF7014), IBA1 (ProteinTech, 10904-1-AP), NLRP3 (Affinity, DF7438), MMP9 (Abmart, TA0220), IL-6 (Abmart, TD6087); cell: iNOS (Affinity, AF0199), CD86(Affinity, DF6332), TLR4 (ProteinTech, 19811-1-AP); JNK (ProteinTech, 24164-1-AP); p38 (ProteinTech, 14064-1-AP); p65 (ProteinTech, 10745-1-AP); p-JNK (Affinity, AF3318); p-p38 (Affinity, AF4001); p-p65 (Affinity, AF2006), TNF-a (Affinity, AF7014), IL-1b (Affinity, AF5103); zebrafish: IL-1b (ProteinTech, 16806-1-AP). b-tubulin (Affinity, AF7018), GAPDH (Affinity, AF7021), b-actin (Affinity, AF7011).

Techniques: Injection

Journal: Advanced science (Weinheim, Baden-Wurttemberg, Germany)

Article Title: Excessive MYC Orchestrates Macrophages induced Chromatin Remodeling to Sustain Micropapillary-Patterned Malignancy in Lung Adenocarcinoma.

doi: 10.1002/advs.202403851

Figure Lengend Snippet: Figure 3. The induction of MP-pattern malignancy relies on macrophages in syngeneic transplant mice with bearing redundant MYC expression tumor. A) Single-cell RNA sequencing (scRNA-seq) data of lung adenocarcinoma from Michael Bartoschek et al. study was utilized. Malignant epithelial cells were selected, and mapping was performed based on bulk RNA-seq data following microdissection, as shown in Figure 1H. Two distinct cell populations were identified: SCISSOR+ cells (MP-subtype) and SCISSOR- cells (AC-subtype). B) Circos plot showing the cell-cell interactions in cell clusters from(A) scRNA-seq after mapping by SCISSOR algorithm. The thickness of each string indicates the number of different interaction pairs colored by cell clusters. C) Representative images of immunohistochemistry of macrophage marker CD68 in tissues micro-array from MAPes cohort. D) Representative images of immunohistochemistry of adenocarcinoma marker TTF1, and multiple immunofluorescences of CD206 (red), CD68 (green) and DAPI (blue) from FFPE slide with mutli-subtypes samples. E) A schematic illustration showing two distinct strategies depleting macrophages in mice with MYC redundant expression tumors. F,G) Representative images of immunohistochemistry of E-cad, TWIST1 and ZEB1 (F), PKP1 and TRPM7 (G). right: quantitative statistics. Five random individual fields were selected from each pathological slide for the assessment of the 12-point IHC score. Scale bars: 50 μm.

Article Snippet: The following primary antibodies were used: Max antibody (CST, #4739), Villin-1 antibody (CST, #55883), CD68 antibody (Proteintech, #28058-1-AP), CD206 antibody (Proteintech, #60143-1-Ig), MHC-II antibody (Santa Cruz, #sc-13556), CD163 antibody (Proteintech, #16646-1- AP), CSF1R antibody (Proteintech, #25949-1-AP), c-Myc antibody (CST, #18583) and FOSL2 antibody (Santa Cruz, #sc-166102).

Techniques: Expressing, RNA Sequencing, Laser Capture Microdissection, Immunohistochemistry, Marker, Microarray

Journal: Frontiers in Nutrition

Article Title: Prevotella histicola Mitigated Estrogen Deficiency-Induced Depression via Gut Microbiota-Dependent Modulation of Inflammation in Ovariectomized Mice

doi: 10.3389/fnut.2021.805465

Figure Lengend Snippet: P. histicola increased the level of BDNF for nerve proliferation. (A) 5-HT1A mRNA level in hippocampus. (B) 5-HT1B mRNA level in hippocampus. (C) DA-1 mRNA level in hippocampus. (D) DA-2 mRNA level in hippocampus. (E) BDNF protein level and the ratio of BDNF to β-tubulin in hippocampus. (F) BDNF mRNA level in hippocampus. (G) Immunohistochemistry of hippocampal proliferation marker Ki-67. Magnification 50 × and 400 ×. Scale bar =20 μm. Data are presented as means ± SEM ( n = 3–4/group). * p < 0.05, ** p < 0.01, ns p > 0.05.

Article Snippet: The sections were incubated with primary antibodies against Iba1 (Novus, NB100-1028, 1:500), ZO-1 (Proteintech, 66378-1-IG, 1:100), Occludin (Proteintech, 27260-1-AP, 1:100), CD68 (Proteintech, 28058-1-AP, 1:100), and Ki-67 (Affinity, AF0198, 1:100) at 4°C overnight.

Techniques: Immunohistochemistry, Marker