Journal: Frontiers in Oncology

Article Title: Differential O - and Glycosphingolipid Glycosylation in Human Pancreatic Adenocarcinoma Cells With Opposite Morphology and Metastatic Behavior

doi: 10.3389/fonc.2020.00732

Figure Lengend Snippet: Immune recognition of glycan structures on PaTu-S and PaTu-T cells. (A) Interaction of immature DCs with PaTu-S and PaTu-T were visualized by fluorescence microscopy. Bar = 100 μm. (B) Binding of immature DCs to PaTu-S and PaTu-T in a cell adhesion assay, in the presence or absence of EGTA. Results are derived from 6 independent experiments using different donors and expressed as average percentage binding ± SEM. (C) Binding of recombinant human galectins Gal-1, Gal-3, and Gal-4 (5 μg/ml) to the PDAC cell lines was measured by flow cytometry. Results are given as average MFI ± SEM of at least 2 independent experiments. (D) Binding of Fc-chimeras of DC-SIGN, MGL, DCIR and Dectin-1 to PaTu-S and PaTu-T cells was measured by flow cytometry. Results are given as average MFI ± SEM of at least 3 independent experiments. * P ≤ 0.05, ** P ≤ 0.01, and *** P ≤ 0.001.

Article Snippet: Mouse anti-human galectin (Gal)-1 , and anti-Tn monoclonal antibodies were kindly provided by Dr. RD Cummings (Boston, USA), goat anti-human Gal-4 was purchased from R&D Systems (Minneapolis, MN) and anti-sialyl Lewis A/CA 19-9 monoclonal antibody was from LifeSpan Biosciences (Seattle, WA).

Techniques: Fluorescence, Microscopy, Binding Assay, Cell Adhesion Assay, Derivative Assay, Recombinant, Flow Cytometry

Journal: bioRxiv

Article Title: mTOR-regulated Mitochondrial Metabolism Limits Mycobacterium-induced Cytotoxicity

doi: 10.1101/2022.01.30.478369

Figure Lengend Snippet: (A - E) Torin1- and DMSO-treated THP-1 macrophages were infected with tdTomato-expressing WT or ⊗ ESX-1 Mm at MOI = 3 and treated with prazosin (PRZ, 20 μM) for 7 hrs. See also Figure S4. (A) Confocal micrographs of Galectin-8 (GAL8) immunofluorescence (green) and Mm fluorescence (magenta) in THP-1 macrophages 7 hpi. GAL8 foci associated with Mm (filled arrowheads) or not associated with Mm (open arrowheads) are shown. Scale bar, 20 μm. (B) Percentage of macrophages with GAL8-associated Mm foci. (C) Percentage of Mm volume associated with GAL8 foci 7 hpi. (D) Percentage of cells that have released cytochrome c 7 hpi. (E) mtor sa1675 5/sa16755 fish and mtor- sufficient siblings were infected with ∼90 fluorescent Mm via the hindbrain ventricle on 2 dpf. On 1 and 2 dpi, fish were injected with ∼3 nL of 300 μM PRZ or 1% DMSO into the hindbrain ventricle. Graph indicates the percentage of animals with mycobacterial cording 3 dpi. Symbols represent values from individual (B, C) imaging fields or (D) individual wells. (B, C) Horizontal lines and (D) bars indicate mean values. (E) Numbers within columns indicate animals per group. Statistical analyses, (B, D) one-way ANOVA with Sidak’s post-test or (E) Fisher’s exact test. * p < 0.05, **p < 0.01, ***p < 0.001, and **** p < 0.0001.

Article Snippet: Cells were then washed twice with PBS and incubated in permeabilization/block (PB) solution (0.1% Triton-X 100, 1% bovine serum albumin in PBS) for 30 minutes at RT, and subsequently stained with goat anti-human galectin-8 antibody (R&D Systems) diluted in PB solution overnight at 4°C.

Techniques: Infection, Expressing, Immunofluorescence, Fluorescence, Injection, Imaging