dylight 649  (Vector Laboratories)

Journal: bioRxiv

Article Title: Epithelial function of the circadian clock gene, Bmal1 , in regulating the mucosa

doi: 10.64898/2026.04.15.718752

Figure Lengend Snippet: ( a )Representative images of Epi Ctrl and Epi ΔBmal1 distal colon stained for UEA-1 (violet) and MALII (yellow) to label fucosylated and sialylated mucins, respectively. Shown are samples collected at ZT0 and ZT12, arrowheads indicate mucus thickness. (Scale bar = 20μm). ( b ) UEA-1+ mucus thickness in the distal colon is significantly higher at ZT12 in Epi Ctrl but lower at both times in Epi ΔBmal1 (**p≤0.01). ( c ) MALII+ mucus thickness in the distal colon is significantly higher at ZT12 in Epi Ctrl (*p≤ 0.05), with a significant increase of thickness in Epi ΔBmal1 at ZT0 (*p<0.05). Note, that Epi ΔBmal1 does not show a time-of-day dependent change in MALII. ( d ) UEA-1+ goblet cell area in distal colon crypts s significantly higher at ZT12 in Epi Ctrl (**p≤0.05). While not significantly different from controls, Epi ΔBmal1 do not show this time-of-day change in UEA-1+ mucus thickness. ( e ) MALII+ goblet cell area in distal colon crypts is significantly increased in Epi ΔBmal1 at ZT0 (*p≤0.05).

Article Snippet: Sections were incubated with biotinylated MAL-II (B-1265-1, Vector Laboratories; 3μg/mL in 1%BSA in PBS) overnight at 4°C, washed in PBS, then incubated with UEA-1 rhodamine (RL-1062-2, Vector Laboratories; 2μg/mL) and streptavidin-FITC (405201, BioLegend; 5μg/mL) in 1% BSA in PBS for 1 hr at room temperature in the dark.

Techniques: Staining

Journal: The American Journal of Pathology

Article Title: Neonatal Thymic Dynamics Influence Autoimmune Pathology by Shaping the Suppressive Potential of Regulatory T Cells

doi: 10.1016/j.ajpath.2025.12.007

Figure Lengend Snippet: Thymic epithelial cell (TEC) subpopulations in the neonatal thymus. A: Representative flow cytometry profiles of CD45 − epithelial cell adhesion molecule (EpCAM) + TECs at postnatal days 1, 3, 5, and 7. B: Total number of TECs in the thymus from the indicated time points. C: Representative flow cytometry profiles of medullary TECs [mTECs; ulex europaeus agglutinin 1 (UEA1) + Ly51 – ] and cortical TECs (cTECs; UEA1 – Ly51 + ) within the CD45 − EpCAM + TEC population from the indicated time points. D: Total number of mTECs ( left panel ) and cTECs ( right panel ) in the thymus from the indicated mice. E: Representative flow cytometry profiles of autoimmune regulator (Aire) expression in mTECs from the indicated time points. F: Frequency ( left panel ) and absolute number ( right panel ) of Aire + mTECs. At least three independent experiments were conducted using neonatal mice. Data are presented as means ± SEM ( B , D , and F ). n = 4 or 5 mice per group ( B , D , and F ). ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001, and ∗∗∗∗ P < 0.0001. Max, maximum; NS, not significant.

Article Snippet: Isolated TECs were stained with the following antibodies: BV421-conjugated anti-mouse CD45 (BD Biosciences, Franklin Lakes, NJ; 30-F11); phosphatidylethanolamine-Cy7–conjugated anti-mouse epithelial cell adhesion molecule (BioLegend; G8.8); Alexa Fluor 647–conjugated anti-mouse Ly-51 (BioLegend; 6C3); DyLight 594–conjugated ulex europaeus agglutinin 1 (UEA1) (Vector Laboratories, Newark, CA; DL-1067-1); and Alexa Fluor 488–conjugated anti-mouse Aire (eBioscience; 5H12).

Techniques: Flow Cytometry, Expressing