Journal: Nature

Article Title: Microglia–neuron crosstalk through Hex–GM2–MGL2 maintains brain homeostasis

doi: 10.1038/s41586-025-09477-y

Figure Lengend Snippet: a , Volcano plot indicating the differentially regulated lipids between Hexb −/− ( n = 6) and Hexb +/− ( n = 3) mice measured by untargeted lipidomics (liquid chromatography–mass spectrometry) at P120. Two-tailed Welch’s t -test was used for statistical testing. b , Spatial MALDI MSI on Hexb −/− and Hexb +/− brains at P0 (upper row), P7 (middle row) and P120 (bottom row). For each indicated ganglioside, ion images representative for three biological replicates are shown. Colour scale represents a visual map of the intensities (in arbitrary units) of the ion images. c , MALDI MSI on the thalamus of Hexb −/− and Hexb +/− mice at P120. For each indicated ganglioside, ion images representative for three biological replicates are shown. Colour scale represents a visual map of the intensities (in arbitrary units) of the ion images. Triangles points to the centromedian and the parafascicular nucleus. d , Hierarchical clustering of selected gangliosides in the indicated CNS specimen. Colour scale indicates the Z -score. e , Experimental scheme. f – h , Left, absolute cytokine and chemokine levels in the supernatant after culturing primary microglia upon overnight ganglioside stimulation. Data are shown as mean ± s.e.m. from four independent replicates. Two-way ANOVA followed by Sidak’s multiple comparison test was used for statistical testing. Right, log 2 [FC] values (colour scale) are shown relative to the unstimulated condition within each genotype. Statistical significance was assessed using one-way ANOVA with Dunnett’s correction. –log 10 [ P ] is encoded in colour intensity (heat map), and cells marked with # indicate P < 0.05. f – h , A comparison of Hexb −/− and Hexb +/+ microglia ( f ), the effect of MGL blockade ( g ) (MGL antibody versus isotype control (ctrl)) and responses to different gangliosides ( h ) (GM1, GM2, GM3). Scale bars, 9 mm ( b ), 2 mm ( c ). Illustrations in e were created using BioRender. Frosch, M. (2025) https://BioRender.com/kdrwxwm .

Article Snippet: For MGL blocking assay, primary microglia were pretreated with anti-MGL antibody (HM1081, 10 μg ml −1 , Hycult Biotech), isotype control IgG (02-9688, 10 μg ml −1 , Thermo Fisher), GalNAc (A2795, 50 mM, Sigma) or EGTA (3054.1, 10 mM, Roth) for 30 min at 37 °C before the ganglioside stimulation.

Techniques: Liquid Chromatography, Mass Spectrometry, Two Tailed Test, Comparison, Control

Journal: Life Metabolism

Article Title: Deletion of hepatic growth hormone receptor contributes to the pathogenesis of lean MAFLD by elevating CD36

doi: 10.1093/lifemeta/loaf037

Figure Lengend Snippet: LGHRKO mice contain excessive hepatic lipids with impaired liver function, similar to DIO mice. (a) Microscopic appearance of the livers in control, DIO, and LGHRKO mice. (a’–c’) H&E-stained liver section at 200× magnification. (d’–f’) H&E-stained liver section at 400× magnification. (g’–i’) Oil Red O-stained liver section at 200× magnification. (j’–l’) Oil Red O-stained liver section at 400× magnification. (b) Hepatic TG content of control, DIO, and LGHRKO mice. (c) Serum AST and ALT levels of control, DIO, and LGHRKO mice. (d) Relative hepatic mRNA levels of lipogenesis genes Acc1 and Fas . (e) Relative hepatic mRNA levels of lipolysis genes Atgl , Hsl , and Mgl . n = 6–8 mice in each group. Data are presented as mean ± SEM.

Article Snippet: Protein lysates (50 μg) were separated by SDS-PAGE, transferred onto nitrocellulose (NC) membranes (#66485; PALL), and blotted with the indicated primary antibodies at 1:1000 dilution: ACC1 (#A15606; ABclonal, RRID: AB_2763012), FAS (#A0461; ABclonal, RRID: AB_2757202), PPARγ (#A0270; ABclonal, RRID: AB_2757083), ATGL (#A6245; ABclonal, RRID: AB_2766852), HSL (#A15686; ABclonal, RRID: AB_2763096), MGL (#A6654; ABclonal, RRID: AB_2767241), CD36 (#A5792, Abclonal, RRID: AB_2766544), β-actin (#AC026, Abclonal, RRID: AB_2768234), and GAPDH (#10494-1-AP; Proteintech, RRID: AB_2263076).

Techniques: Control, Staining

Journal: Immunity

Article Title: Mitochondrial Membrane Potential Regulates Nuclear Gene Expression in Macrophages Exposed to Prostaglandin E2

doi: 10.1016/j.immuni.2018.10.011

Figure Lengend Snippet: (A-B) RNA-seq analysis of gene expression in bone marrow macrophages (BMMs) stimulated for 6 h with: IL-4 (M(IL-4)) or PGE2 (M(−)+PGE2)(A) or M(IL-4) or M(IL-4)+PGE2 (B), compared to no stimulus controls (M(−)) (A,B). (C) Expression of CD301 and RELMα proteins, as measured by flow cytometry, in BMMs stimulated for 24 h with indicated treatments. MFI = mean fluorescence intensity. (D) Western blot of phosphorylated STAT6 (P-STAT6) at indicated times in BMMs treated as indicated. ACTB, actin loading control. (E-F) Extracellular flux analysis (EFA) of oxygen consumption rates (OCR) (E) or extracellular acidification rates (ECAR) (F) at 24 hr post stimulation. For EFA, BMMs were sequentially treated with oligomycin (Oligo), FCCP and rotenone plus antimycin A (R/A) as indicated. (G) 13C-Glucose LC-MS trace into TCA cycle metabolites from cells stimulated for 24 h with indicated treatments. (H-K) ATP concentration (H), cytoplasmic ROS (I), mitochondrial ROS (J), and Δψm as measured by TMRM incorporation (K), in BMMs stimulated as indicated for 24 h. (A-B) Significant (adjusted p value < 0.1) > 2 fold up or down regulated genes based on 3 biological replicates denoted by *. (C) Mean ± SEM from 5 biological replicates (p***<0.0005, p****<0.0001). (G) Mean ± SEM from 3 biological replicates, normalized to M(−) (p**<0.005). (H-K) Mean ± SEM from 5 biological replicates (p*<0.05, p***<0.0005, p****<0.0001). Data are representative of 2 (D) or 3 (C, E, F, H-K) independent experiments. See also Figure S1.

Article Snippet: Pathway enrichment analysis was performed using Ingenuity Pathway Analysis (Qiagen), which makes predictions using pair-wise comparisons between conditions, incorporating in the analysis the statistically significant changes in expression within each of those comparisons. . Flow cytometry Used fluorochrome-conjugate monoclonal antibodies included: CD301 (Milteny Biotech, clone: REA687), CD11b (Biolegend, clone: M1/70), F4/80 (Biozol, clone: BM8), TIM4 (BioLegend, clone: F31–5G3), CD45.1 (BioLegend, clone: A20).

Techniques: RNA Sequencing, Gene Expression, Expressing, Flow Cytometry, Fluorescence, Western Blot, Control, Liquid Chromatography with Mass Spectroscopy, Concentration Assay

Journal: Scientific Reports

Article Title: Monoacylglycerol Lipase: A Novel Potential Therapeutic Target and Prognostic Indicator for Hepatocellular Carcinoma

doi: 10.1038/srep35784

Figure Lengend Snippet: Methylation special primers were detected with MSP in all HCC cell lines but in L02 by MSP. Unmethylation special primers were only detected in L02. After treatment by DAC, methylation special primers decreased in HCC cell lines while unmethylation special primers increased ( A ). In HepG2 and SMMC-7721 cells ( B – D ), expression of LATS1 protein was suppressed. Demethylation of LATS1 promoter by DAC and knock-in LATS1 gene by plasmid transfection elevated LATS1 protein levels, which confirmed our findings that methylation of LATS1 promoter induced lost-expression of LATS1 protein in HCC. In HepG2 and SMMC-7721 cell lines, regulation of LATS1 with these methods induced significant changes in downstream signals YAP protein levels in a negatively correlative manner and pYAP protein levels in a positively correlative manner. Finally, due to methylation of LATS1 promoter and lost expression of LATS1, YAP protein could not be phosphorylated and detained in cytoplasm effectively. Excess YAP imported into nucleus and induced overexpression of MAGL.

Article Snippet: After transfer membranes were blocked with 5% nonfat milk, incubated with polyclonal antibodies for MAGL, LATS1, YAP1 or pYAP1 (Abcam, USA) respectively, and a monoclonal antibody for β-actin (Boster, China) was used as a loading control.

Techniques: Methylation, Expressing, Knock-In, Plasmid Preparation, Transfection, Over Expression