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: Biology of Reproduction

Article Title: Sialylation Facilitates the Maturation of Mammalian Sperm and Affects Its Survival in Female Uterus 1

doi: 10.1095/biolreprod.115.137810

Figure Lengend Snippet: Sialome profile of sperm from testis to ejaculated sperm. A ) Summary of sperm sialylation status during different developmental stages by lectin histology and anti-Neu5Gc immunohistochemical staining. Except for the presence of Sia α2-3 on spermatocytes, incremental sialylation of other types from spermatogonia to ejaculated sperm. B ) Sia α2-3 presence on spermatocytes and Sia α2-3/6 and Neu5Gc presence on sperm in cauda of epididymis. Frozen sections of testis and epididymis of WT male mice were fixed with acetone; incubated with SNA, MAL-II, and anti-Neu5Gc antibody as primary antibody and HRP-anti-streptavidin and HRP-anti-chicken antibody as secondary antibody; and finally developed with AEC substrate chromogen. Typical samples were chosen to show the presence of Sia α2-3 presence on spermatocytes and Sia α2-3/6 and Neu5Gc presence on sperm in cauda of epididymis (black arrows). Bars = 50 μm (top) and 500 μm (bottom). C ) Quantification of incremental sialome of Sia α2-3/6 and Neu5Gc on sperm from caput to cauda of the epididymis. Sperm samples collected from caput to cauda of epididymis were fixed by PFA and detected for Sia α2-3/6 lectin staining and anti-Neu5Gc antibodies staining via flow cytometry. Sialylation levels increased from the caput to cauda of epididymis in male mice.

Article Snippet: SNA, MAL-II, and anti-Neu5Gc antibody (Sialix, Inc.) were used at a concentration of 1:500 for 1 h at RT, and then HRP-anti-streptavidin and HRP-anti-chicken antibody were used at 1:500 for 30 min at RT.

Techniques: Immunohistochemical staining, Staining, Incubation, Flow Cytometry

Journal: Biology of Reproduction

Article Title: Sialylation Facilitates the Maturation of Mammalian Sperm and Affects Its Survival in Female Uterus 1

doi: 10.1095/biolreprod.115.137810

Figure Lengend Snippet: Detection of sialoglycoprotein in epididymal fluid from different epididymal segments. A ) Contents and semiquantification of the regional epididymal fluid on reduced SDS-PAGE gel by Coomassie staining, loading, 10 μg protein/lane. Sialoglycoproteins were found throughout the epididymis; the amount and composition varied between fluids from different epididymal segments. B ) Sia α2-3 glycoproteins in regional epididymal fluids. Sia α2-3 glycoproteins in the samples were detected by MAL-II-specific binding (loading: HSA, 100 ng; others, 10 μg protein/lane). HSA was used as positive control in the first lane. C ) Sia α2-6 glycoproteins in the regional epididymal fluids. Sia α2-6 glycoproteins in the samples were detected by SNA-specific binding (loading: HSA, 100 ng; others, 10 μg protein/lane). HSA was used as positive control in the first lane. D ) Neu5Gc-glycoproteins in regional epididymal fluids. Neu5Gc-glycoproteins in the samples were detected by anti-Neu5Gc antibody (loading: WT mouse sera, 100 μg; others, 10 μg protein/lane). Mouse serum was used as positive control in the first lane.

Article Snippet: SNA, MAL-II, and anti-Neu5Gc antibody (Sialix, Inc.) were used at a concentration of 1:500 for 1 h at RT, and then HRP-anti-streptavidin and HRP-anti-chicken antibody were used at 1:500 for 30 min at RT.

Techniques: SDS Page, Staining, Binding Assay, Positive Control