anti cd25 antibody  (MedChemExpress)

Journal: bioRxiv

Article Title: Single recipient cell tracking of tellurium-labeled extracellular vesicle proteomes (TeLEV) identifies EV-driven immunomodulation

doi: 10.1101/2025.11.01.685872

Figure Lengend Snippet: a, t-SNE projection of PBMC recipient cells across 16 conditions (PBS-HAT control plus 15 EV sources, in total 2,977,094 cells), colored according to cell type. Numerical annotations indicate the relative proportions of each cell lineage. EVs shown in were isolated from 25 mL of cell-conditioned medium (from 25M primary CLL cells or cell lines over 48 hours). The EV uptake conditions were normalized according to the input volume of cell-conditioned media (25 mL, 25M cells over 48 hours), and EV counts and buffer volumes are reported in Supplementary Table 2. b, t-SNE projection of PBMC recipients across 16 conditions showing TeLEV signal. c, Fold changes in cell type proportions across conditions, estimated using a Poisson generalized linear mixed model that accounts for confounding factors (Methods). Dot size represents the probability of change, measured as the local true sign rate (LTSR). d, t-SNE projection of myeloid cells across 16 conditions (276, 867 cells), colored according to subtypes. e, t-SNE plots showing expression of monocytic and DC lineage markers and interleukin receptors on myeloid cells. f, Two-dimensional density distributions of myeloid cells in t-SNE embedding space, stratified by conditions. g, A cell type–color–coded ternary plot depicting expression levels of CD25, CD123, and CD127. Circle size reflects the average TeLEV signal level within each cell type. Colors correspond to those in the legend shown in (a)

Article Snippet: Anti-CD25 antibody (Basiliximab, MedChemExpress, #HY-10885) was used at 10 μg/mL and compared to an isotype control (Invitrogen, #MA5-55090).

Techniques: Control, Isolation, Expressing

Journal: bioRxiv

Article Title: Single recipient cell tracking of tellurium-labeled extracellular vesicle proteomes (TeLEV) identifies EV-driven immunomodulation

doi: 10.1101/2025.11.01.685872

Figure Lengend Snippet: a, t-SNE projection of PBMC recipient cells across 11 time points (0-48h, in total 1,512,100 cells) of EV uptake, colored according to cell type. Numerical annotations indicate the relative proportions of each cell lineage. Primary MBC-EVs and secreted proteins shown in were isolated from 4,2 mL of cell-conditioned medium (from 4.2M primary CLL cells over 48 hours), resulting in 2.7 x 10 9 EVs used per time point condition. b, t-SNE plots showing TeLEV signal, CD25, CD127, and CD123 expression at selected time points (columns). c, Stacked bar plot depicting changes in cell type composition over 48h. Colors correspond to those in the legend shown in (a). d, Comparison of TeLEV signal distribution across major cell types and their subtypes at 16 h, with colors representing the average TeLEV signal level in each population. Bars above the plots show the relative proportions of each population. White lines denote the median, edges the IQR, and whiskers either 1.5 × IQR or minima/maxima (if no point exceeded 1.5 × IQR; minima/maxima are indicated by the violin plot range). e, Comparison of TeLEV signal, CD25, CD123, and CD127 distributions within the Monocyte population (including Non-classical Monocytes, Intermediate Monocytes, Classical Monocytes, IL2R/IL7R + Monocytes, and CD197 + IL2R/IL7R + Monocytes) over 48 h. Statistical significance across all time points for each marker was evaluated using the Kruskal-Wallis test. Pairwise comparisons between 00 h and 00 h 15 min were performed using a one-sided Mann–Whitney U test. p-values < 0.05 were considered significant and are indicated in the figures as follows: * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001. f, Line plots depicting the mean expression levels of TeLEV signal, CD25, CD123, and CD127 within the Monocytes during the first 2 hours of EV uptake. Error bars represent mean ± SEM. g, The same analysis as in (e) for mDCs (left) and pDCs (right). h, Live-cell immunocytochemistry of purified monocytes exposed to increasing doses of Ramos-derived EVs. Representative frames from day 2 and day 7 are shown. CD25 (Alexa Fluor 488; green) and Cytotox NIR (blue; loss of membrane integrity) are displayed after automated single-cell segmentation (Incucyte built-in). Cells were seeded at 25,000 per well and imaged every 2 h for 7 days using an Incucyte SX5 (20× objective). Scale bar, 200 µm. EVs and secreted proteins shown in were isolated from 1.5 mL of cell-conditioned medium (from 1.5M Ramos cells over 48 hours), resulting in 9 x 10 8 EVs used per EV condition (3X condition, ). i, Single-cell quantification of CD25 (Alexa Fluor 488) fluorescence over 7 days corresponding to (h). Values are unnormalized arbitrary units; data shown as mean ± SEM; n = 4 fields of view. j, Flow cytometric contour plots of purified monocytes after 16 h of EV exposure across a dose titration (see Figure legend ). CD25 (APC), CD123 (PE), and CD127 (FITC) were measured on a Miltenyi MACSQuant X with compensation using BioLegend compensation beads. DAPI was used as a surface-flow viability dye. k, Live-cell immunocytochemistry of EV-exposed monocytes stimulated with cytokines. Representative images from day 7 are shown for IL-2 (20 ng mL −1 ), IL-3 (5 ng mL −1 ), or, IL-7 (25 ng mL −1 ), or IL-2 + IL-3, IL-2 + IL-7, IL-3 + IL-7, or IL-2 + IL-3 + IL-7 in RPMI with serum. CD25 (Alexa Fluor 488; green) and Cytotox NIR (blue) are shown after automated segmentation and background subtraction. Imaging on Incucyte SX5 (20×, 2 h intervals). Scale bar, 200 µm. l, Immunogold transmission electron microscopy of Ramos-EVs labeled for IL-3, CD123, and CD127 alongside an isotype control. Scale bar, 100 nm. m, Intracellular phospho-flow cytometry of purified monocytes incubated with EVs, showing total STAT5 (Alexa Fluor 488), total STAT3 (PE), pSTAT5 (PE-Cy7), and pSTAT3 (BV421). EV exposure was terminated by immediate formaldehyde fixation after 16 h prior to staining. Cleaved caspase was included as an apoptosis marker. Acquisition on a Miltenyi MACSQuant X with bead-based compensation of all markers. n, Live-cell immunocytochemistry of EV-exposed healthy-donor PBMCs stimulated for 2 days with IL-2 (20 ng mL −1 ), IL-3 (5 ng mL −1 ), or IL-7 (25 ng mL −1 ) in RPMI with serum. Representative images are shown (Incucyte SX5, 20×, 2 h intervals). Scale bar, 200 µm. o, Live-cell immunocytochemistry of EV-exposed healthy-donor PBMCs treated for 7 days with a STAT5 degrader (AK-2292) or anti-CD25 (Basiliximab), compared with DMSO and isotype controls. CD25 (Alexa Fluor 488; green), CD127 (PE; red), and Cytotox NIR (blue) are shown after automated segmentation and background subtraction. Cells were seeded at 50,000 per well and imaged on an Incucyte SX5 (20×, every 2 h). Scale bar, 200 µm. p, Flow-cytometric contour plots of healthy-donor PBMCs after 48 h of EV exposure across a dose titration (1× = 3×10 8 EVs per 10 5 cells), showing CD25 (APC), CD123 (PE), and CD127 (FITC). Acquisition and compensation as in (m).

Article Snippet: Anti-CD25 antibody (Basiliximab, MedChemExpress, #HY-10885) was used at 10 μg/mL and compared to an isotype control (Invitrogen, #MA5-55090).

Techniques: Isolation, Expressing, Comparison, Marker, MANN-WHITNEY, Immunocytochemistry, Purification, Derivative Assay, Membrane, Fluorescence, Titration, Imaging, Transmission Assay, Electron Microscopy, Labeling, Control, Flow Cytometry, Incubation, Staining

Journal: bioRxiv

Article Title: Single recipient cell tracking of tellurium-labeled extracellular vesicle proteomes (TeLEV) identifies EV-driven immunomodulation

doi: 10.1101/2025.11.01.685872

Figure Lengend Snippet: a, t-SNE projection of 16 conditions (in total 3,590,520 cells) coloured according to cell type. Numerical annotations indicate the relative proportions of each cell lineage. MBC-EVs shown in were isolated from 8 mL of cell-conditioned medium (from 8M Ramos cells over 48 hours), resulting in 5 x 10 9 EVs used per EV condition. b, Stacked bar plot depicting changes in cell type composition. Colors correspond to those in the legend shown in (a). c, Comparison of TeLEV signal distribution across major cell types and their subtypes. Shown are EVs (217,211 cells), STAT5d + EVs (226,884 cells), and STAT3d + EVs (206,018 cells) conditions, with colors representing the average TeLEV signal level in each population. Bars above the plots show the relative proportions of each population in the corresponding condition (left). White lines denote the median, edges the IQR and whiskers either 1.5 × IQR or minima/maxima (if no point exceeded 1.5 × IQR; minima/maxima are indicated by the violin plot range). t-SNE projection of cells for each condition showing TeLEV signal (middle). Two-dimensional density distributions of cells in t-SNE embedding space, stratified by conditions (right). d, Comparison of TeLEV signal distribution across three conditions shown in (c). Pairwise comparisons were performed using a two-sided Mann–Whitney U test. **** p < 0.0001. e, t-SNE projection of myeloid population of 16 conditions (865,413 cells), colored according to subtypes. f, t-SNE plots showing expression of monocytic and DC lineage markers and interleukin receptors on myeloid cells. g-k, t-SNE plots showing CD25, CD127, CD123, BCL-2, and PD-L1 expressions across the indicated conditions. l, Comparison of PD-L1 expression distribution across the indicated conditions in Monocytes, mo-derived DCs, mDCs, and pDCs. For each cell type, statistical significance across all conditions was evaluated using the Kruskal-Wallis test.**** p < 0.0001. m, Same as in (l) for BCL-2. n-p, Comparison of CD25 (n), Ki-67 (o), and CD69 (p) expression distributions across major cell types and their subtypes, with colors representing the average respective marker expression level in each population. Shown are IL-2 (238,611 cells) and IL-2 + EVs (207,300 cells) conditions.

Article Snippet: Anti-CD25 antibody (Basiliximab, MedChemExpress, #HY-10885) was used at 10 μg/mL and compared to an isotype control (Invitrogen, #MA5-55090).

Techniques: Isolation, Comparison, MANN-WHITNEY, Expressing, Derivative Assay, Marker

Journal: Cell Reports Medicine

Article Title: A multiomics analysis-assisted deep learning model identifies a macrophage-oriented module as a potential therapeutic target in colorectal cancer

doi: 10.1016/j.xcrm.2024.101399

Figure Lengend Snippet:

Article Snippet: Mice were treated with diphtheria toxin (MCE, #HY-108851, USA) or 50 mg/kg 5-Fluorouracil (5FU) (MCE, #HY-90006, USA) through intraperitoneal injection for 2 weeks before sacrificed, and the injections were performed every 3 days.

Techniques: Purification, Recombinant, Protease Inhibitor, Red Blood Cell Lysis, Polymer, Blocking Assay, Modification, H&E Stain, Immunohistochemical staining, Sequencing, RNA Sequencing Assay, Software