Journal: Frontiers in Pharmacology

Article Title: Mycobacterium tuberculosis infection drives osteoclast overactivation via α2,3-Sialylation to promote pathological bone destruction

doi: 10.3389/fphar.2026.1738896

Figure Lengend Snippet: α2,3-sialylation is required for BCG-induced osteoclast differentiation and activity (A) Comparison of differentially expressed genes (DEGs) between BCG-infected osteoclasts (RB) and uninfected controls (R). (n = 3 biological replicates per group, differential expression was defined as |FoldChange| > 2 and padj <0.05). (B) Volcano plot of DEGs highlighting genes related to sialic acid biosynthesis. (C) KEGG pathway enrichment analysis of upregulated DEGs in RB cells. (D,E) Immunofluorescence staining of α2,3-SA in mouse calvarial sections using MAL II lectin, with quantification of α2,3-SA fluorescence intensity (n = 5). Intensity density was normalized to the PBS group mean. (F) In vitro osteoclasts subjected to MAL II lectin staining and TRAP staining, with or without sialidase treatment to enzymatically remove α2,3-SA. (G) Quantification of α2,3-SA fluorescence intensity in cultured osteoclasts (n = 5). Intensity density was measured in cellular ROIs after background subtraction and normalized to the RANKL group mean. (H) Quantification of TRAP + multinucleated cells (≥3 nuclei) per field (randomly selected fields, fixed magnification) in vitro (n = 5). (I) mRNA expression levels of osteoclast differentiation markers ( Fos, Mmp9, Nfatc1, and Ocstamp ) in osteoclasts (n = 3). Data are presented as mean ± SD. Statistical significance was determined by two-tailed unpaired Student’s t-test for two-group comparisons (E) and one-way ANOVA followed by Tukey’s post hoc test for three-group comparisons (G–I) .

Article Snippet: Biotin MAL-II , Vector laboratories , Cat# B-1265-1.

Techniques: Activity Assay, Comparison, Infection, Quantitative Proteomics, Immunofluorescence, Staining, Fluorescence, In Vitro, Cell Culture, Expressing, Two Tailed Test

Journal:

Article Title: PDX-1 haploinsufficiency limits the compensatory islet hyperplasia that occurs in response to insulin resistance

doi: 10.1172/JCI200421845

Figure Lengend Snippet: Immunostaining for pancreatic ductal marker, and markers for cell death and neogenesis. (A and B) Representative sections of pancreas from different genotypes, showing islets stained for PCNA (A) and caspase-3 (B). Note the magnified image for TKO. (C) Consecutive sections of pancreas from IR/IRS-1 mice immunostained for PCNA (left panels) or costained for insulin (green) and DBA-lectin immunohistochemistry for duct-specific glycoconjugates (red) (upper right panel). The lower right panels show 2 magnified images (×60) of islet cells from IR/IRS-1 mice; the arrows point to cells positive for PCNA (purple chromogen) that also stain positive for nuclear β-catenin (blue chromogen). (D) Pancreas sections from WT, IR/IRS-1, PDX-1 heterozygous, IRS-1/PDX-1, and TKO mice costained for E-cadherin (purple) and β-catenin (orange) as described in Methods. Scale bars: 50 μm.

Article Snippet: Primary antibodies included guinea pig anti-insulin (Linco Research Inc.) or sheep anti-insulin (The Binding Site Ltd.), rabbit anti-somatostatin (DAKO), mouse anti-glucagon (Sigma-Aldrich), rabbit anti-Glut2 (a gift from B. Thorens, University of Lausanne, Lausanne, Switzerland), rabbit anti-synaptophysin (DAKO), mouse anti-PCNA (DAKO), β-catenin (BD Biosciences — Pharmingen), E-cadherin (Valeant Pharmaceuticals), rabbit anti–caspase-3, and Texas red–conjugated lectin (EY Laboratories Inc.).

Techniques: Immunostaining, Marker, Staining, Immunohistochemistry

Journal: BioMed Research International

Article Title: Sialic Acid Expression in the Mosquito Aedes aegypti and Its Possible Role in Dengue Virus-Vector Interactions

doi: 10.1155/2015/504187

Figure Lengend Snippet: DENV interaction with Ae. aegypti SG. (a) DENV interaction with Ae. aegypti SGs. SGs from Ae. aegypti were incubated with DENV and stained with anti-DENV E antibody and rhodamine-coupled anti-IgG antibody. (b) DENV-SG competence assays using ConA, LCH, and SNA lectins, which were added to SG before incubation with DENV. The interaction with DENV was blocked when DENV was incubated in the presence of lectins that recognized Sia. With LCH and ConA lectins, the magnification = 10x and with SNA lectin = 20x. Scale bar = 10 μ m. (c) DENV-SG interaction in the absence or presence of sialidase. SGs were untreated or pretreated with C. perfringens sialidase for 30 min before adding DENV. The DENV-SG interactions in the presence of Sia competitors, fetuin (1 mM) and free Sia (200 nM), are also shown, where the DENV-SG interaction was blocked. (d) DENV-SG interaction in SGs pretreated with trypsin for 5, 15, or 30 min before adding DENV. There was a decrease in the DENV-SG interaction after 15 min, and it was lost completely at 30 min. Scale bar = 10 μ m. Blue: nuclei stained with DAPI. Red: DENV stained with an antibody against viral protein E and a secondary antibody coupled to rhodamine. Green: (FITC) SNA lectin interaction.

Article Snippet: Ae. aegypti SGs and midguts were placed on slides and fixed, and the tissues were then blocked with 2% bovine serum albumin (BSA) for 30 min at RT, washed with PBS for 5 min, and immersed in PBS-Triton X-100 (0.2%) for 10 min. Next, they were washed with PBS-Ca 2+ (1 mM) for 10 min and incubated with different biotin-conjugated lectins, that is, Maackia amurensis lectin (MAA), Sambucus nigra agglutinin (SNA), or Lens culinaris hemagglutinin (LCH) (EY Laboratories, Inc, USA) at 1 : 100 dilutions for 2 h at 37°C.

Techniques: Incubation, Staining

Journal: bioRxiv

Article Title: Nontypeable Haemophilus influenzae redox recycling of protein thiols promotes resistance to oxidative killing and bacterial survival in biofilms in a smoke related infection model

doi: 10.1101/2021.08.25.457736

Figure Lengend Snippet: Immunofluorescent imaging and RT-qPCR analysis of NTHi biofilms in the smoke exposed murine lung. A) Immunofluorescent staining of parent strain infected mouse lung sections. Lung sections are representative of 24 and 48 hours post infection. Nuclei were stained with DAPI (blue) and bacteria were stained with anti-NTHi polyclonal antibodies conjugated to Alexa 488 (green). NTHi biofilm components were stained using the Maackia amurensis lectin (MAA) conjugated to Texas-Red, which is specific for neu5Ac α( , ) galactose linkages (red). Images are taken at 90X magnification. Scale bars represent 10 μm. B) RT-qPCR analysis of mRNA isolated from infected mouse lung of biofilm associated genes, in comparison to omp26 housekeeping gene. Samples were collected from 24 hours post infection. All samples were run in triplicate. Mean +/- SD N=4-6.

Article Snippet: Maackia amurensis lectin (MAA) Texas Red conjugated specific for Neu5Ac α( , ) galactose (EY Laboratories, San Mateo, CA) was diluted to a final concentration of 100 μg/ml in 0.01 M phosphate, 0.15 M NaCl, 0.05 M sodium azide buffer according to manufacturer’s instructions.

Techniques: Imaging, Quantitative RT-PCR, Staining, Infection, Isolation