Journal: Research and Practice in Thrombosis and Haemostasis

Article Title: A red blood cell–based antigen delivery system to facilitate T cell epitope presentation to promote peripheral tolerance to ADAMT S13 in immune-mediated thrombotic thrombocytopenic purpura

doi: 10.1016/j.rpth.2026.103387

Figure Lengend Snippet: Monitoring of peptide binding to red blood cells (RBCs). (A) Schematic representation of the peptides used in this study. The TAT-CUB2 peptide consists of a cell-penetrating TAT peptide (dark blue), a short glycine linker (gray), and a CUB2-derived sequence (teal), with the major histocompatibility complex class II–presented core peptide underlined. The CUB2 control peptide lacks the TAT sequence. Peptides used for flow cytometry also include a biotin label for detection (between brackets). (B) Detection of peptide binding to RBCs via flow cytometry. RBCs were incubated with 10, 20, or 30 μM TAT-CUB2-biotin (blue) or CUB2-biotin control peptides (red). RBCs that were not incubated with peptide but otherwise followed the same procedure are represented in gray. Streptavidin–phycoerythrin (PE)–cyanine 7 (Cy7) fluorescence intensity (biotin detection) is represented on the x-axis, and the cell count (number of events) is represented on the y-axis. (C) Detection of peptide binding to RBCs via ImageStream. Representative brightfield and fluorescence images of RBCs after incubation with 20 μM TAT-CUB2-biotin (top) or CUB2-biotin control peptide (bottom). Peptides were visualized by their biotin label via streptavidin coupled to AF488 (green) and PE-Cy7 (pink); the membrane protein glycophorin A is stained by PE-conjugated anti-glycophorin A (orange). TAT, transactivator of transcription.

Article Snippet: PE-conjugated anti-human glycophorin A (R&D Systems) was used to stain the RBC membrane.

Techniques: Binding Assay, Derivative Assay, Sequencing, Immunopeptidomics, Control, Flow Cytometry, Incubation, Fluorescence, Cell Characterization, Membrane, Staining

Journal: Blood Science

Article Title: Large-scale proteomic and phosphoproteomic analysis of erythroid enucleation and maturation

doi: 10.1097/BS9.0000000000000263

Figure Lengend Snippet: Differential analysis of the whole proteome and phosphoproteome across stages of human erythroid enucleation and maturation. A and B, Scatter plots of protein abundance between 2 adjacent erythroid differentiation stages, where the color of each point represents the magnitude of the GFOLD value. C and D, Scatter plots depicting changes in phosphorylated protein abundance between 2 adjacent erythroid differentiation stages, where point color indicates the magnitude of GFOLD values. E and F, GO enrichment results for differentially phosphorylated proteins in the comparison of Ebasts vs Retics (E) or Retics vs RBCs (F). G, Flow cytometric detection of GPA-positive rate in erythroid differentiated cells stained with PE-CD235a after treatment with different apoptosis inducers, with the percentage in red font indicating the GPA-positive rate. H, Flow cytometric detection of enucleation rate of in vitro induced erythroid differentiated cells treated with different apoptosis inducers, which were stained with Hoechst 33342, and enucleated cells were Hoechst 33342-negative. Statistics are shown as the mean ± SD. Comparisons between 2 groups were evaluated with a 2-tailed t test, and comparisons among multiple groups were evaluated with one-way ANOVA. * p < 0.050; ** p < 0.010; *** p < .001. DMSO = dimethyl sulfoxide, GFOLD = generalized fold change, RBC = red blood cell.

Article Snippet: Antibodies used for flow cytometry included PE‐CD34, PE-CD235a (GPA), and APC‐α4 integrin (Miltenyi Biotec, Bergisch Gladbach, Germany); PE‐Cy7‐Annexin V (eBioscience, San Diego, California); and human band 3, which was generated in our laboratory and labeled with FITC.

Techniques: Quantitative Proteomics, Comparison, Staining, In Vitro