Journal: mSphere

Article Title: Use of a human immortalized microglia cell line to study recognition, phagocytosis, and intracellular survival of Cryptococcus neoformans

doi: 10.1128/msphere.00838-25

Figure Lengend Snippet: Phagosome membrane damage occurs in cryptococcal-containing phagosomes. ( A ) Representative image showing a cryptococcal-containing phagosome positive for Gal-3 association. Gal-3 is widely used as a reporter of phagosomal membrane damage. These images were taken at 100×, with blue showing the nuclei of the C20 cells, green showing intracellular KN99α, and yellow showing Gal-3. The scale bar is 10 μm. ( B ) Percentage of fungal-containing phagosomes positive for Gal-3 association 24 h after engulfment was analyzed by immunofluorescence as shown in panel A . Both C. neoformans (KN99α)- and S. cerevisiae (BY4741)-containing phagosomes were tested for membrane damage. mCherry-expressing fungi were either unopsonized (unop.) or opsonized (serum) and incubated with microglia at an MOI of 20:1 for 3 h. Wells were washed three times with DPBS to remove extracellular fungi, and microglia were incubated for an additional 24 h then assessed for membrane damage via Gal-3 staining. Values represent the mean ± SEM from three biological replicates. For all experiments, significance was determined using one-way ANOVA with multiple comparisons (Brown-Forsythe and Welch’s corrected), *, P < 0.05; **, P < 0.005; ***, P < 0.0005; ****, P < 0.0001.

Article Snippet: Cells were immunolabeled in 1% goat serum at 4°C overnight with an antibody against Gal-3 (rat; M3/38: sc23938, Santa Cruz Biotechnology; 1:100 concentration), LAMP1 (mouse; Developmental Studies Hybridoma Bank, H4A3; 1:500 concentration), EEA1 (rabbit; Abcam, ab2900; 1:100 concentration), or vATPase (mouse; Santa Cruz Biotechnology, D-11; 1:500 concentration).

Techniques: Membrane, Immunofluorescence, Expressing, Incubation, Staining

Journal: European Radiology Experimental

Article Title: An endovascular porcine model of abdominal aortic aneurysm for interventional radiology research

doi: 10.1186/s41747-025-00673-z

Figure Lengend Snippet: Western blot analysis of inflammatory and structural protein expression. a Representative western blot showing Galectin-3/Mac2 (Gal-3) expression in aortic tissue at 2 weeks (2 W) and 4 weeks (4 W) post-intervention versus control. b Quantification of Gal-3 levels reveals significant upregulation at 2 W (46.55 ± 7.31%) and 4 W (87.22 ± 11.14%) compared to control (18.86 ± 10.37%; p < 0.001). c WB showing α-smooth-muscle-actin (α-SMA) expression in the same groups. d Quantification shows reduced α-SMA expression at 2 W (52.79 ± 14.99%) and 4 W (39.16 ± 17.52%) versus control (80.94 ± 14.26%; p = 0.005 and p < 0.001, respectively). Individual values: n = 4 for 2 W (dots); n = 4 for 4 W (squares); n = 4 for controls (triangles). Statistical significance: ** p < 0.01, *** p < 0.001. 2 W, Two weeks; 4 W, Four weeks; α-SMA, α-smooth-muscle-actin; Gal-3, Galectin-3

Article Snippet: Macrophage content was evaluated using Galectin-3 (Gal-3) antibody (#14979-1-AP, 1:100; Proteintech Group Inc.).

Techniques: Western Blot, Expressing, Control

Journal: European Radiology Experimental

Article Title: An endovascular porcine model of abdominal aortic aneurysm for interventional radiology research

doi: 10.1186/s41747-025-00673-z

Figure Lengend Snippet: Immunofluorescence analysis of macrophage infiltration and vascular smooth muscle cell loss. a Representative images of aortic tissue stained for Galectin-3 (Gal-3, magenta), α-smooth-muscle-actin (α-SMA; magenta), and DAPI (blue), in control, 2 weeks (2 W), and 4 weeks (4 W) post-intervention samples. Scale bars: 500 µm. b Gal-3-positive area increased significantly at 2 W (15.12 ± 3.88%) and 4 W (16.65 ± 5.27%) versus control (0.66 ± 0.27%; p = 0.012, and p = 0.021, respectively). c α-SMA-positive area decreased at 2 W (3.09 ± 0.70%) and 4 W (2.04 ± 1.43%) versus control (35.18 ± 5.15%; p = 0.003 for both comparisons). Quantified with Keyence Hybrid Cell Count (autothreshold; Version 2.1.1, Keyence Corp.), values signify positive area fraction. Individual values: n = 4 for 2 W (dots); n = 4 for 4 W (squares); n = 4 for controls (triangles). Statistical significance: * p < 0.05, ** p < 0.01. 2 W, Two weeks; 4 W, Four weeks; α-SMA, α-smooth-muscle-actin; Gal-3, Galectin-3

Article Snippet: Macrophage content was evaluated using Galectin-3 (Gal-3) antibody (#14979-1-AP, 1:100; Proteintech Group Inc.).

Techniques: Immunofluorescence, Staining, Control, Cell Characterization

Journal: mBio

Article Title: Loss of LafB activity reverses daptomycin resistance in E. faecium

doi: 10.1128/mbio.00715-25

Figure Lengend Snippet: TLC analysis of glycolipids in total membrane lipid extracts from E. faecium . Glycolipid profiles of E. faecium SM1, SM1 lafB T577C, SM1Δ lafB , and SM1 Δ lafB /pAT28:: lafB mutants were compared to WT E. faecium SM1. Purified lipid standards included Gal₂DAG [1,2-diacyl-3-O-(α-D-galactosyl(1→6)-β-D-galactosyl)-sn-glycerol] and Glc ₁ DAG [1,2-diacyl-3-O-(α-D-glucopyranosyl)-sn-glycerol] (Avanti Polar Lipids). Glycolipid species Glc ₁ DAG, Glc ₂ DAG, and Gal ₂ DAG were identified based on migration relative to standards. Additional bands representing unknown lipid species are also indicated. The absence of glycolipid bands in the Δ lafB mutant and their restoration in the complemented strain demonstrates the role of lafB in glycolipid biosynthesis. Data are representative of three independent experiments.

Article Snippet: Approximately 5 μL of each of the two standards (Gal 2 DAG: 1,2-diacyl-3- O -(α- d -galactosyl1-6)-β- d -galactosyl- sn -glycerol Avanti Polar Lipids 840524P, Glc 1 DAG: 1,2-diacyl-3-O-(α-D-glucopyranosyl)-sn-glycerol Avanti Polar Lipids 840522P) and ~15 μL of each sample was spotted on the baseline of a Silica gel 60 TLC plate.

Techniques: Membrane, Purification, Migration, Mutagenesis