Journal: bioRxiv

Article Title: Translational Opportunity of Engineered IFNγ-eEVs Through Targeted Inhibition of JAK/STAT1 Signaling, Mimicking IVIg Therapy

doi: 10.64898/2026.04.29.721601

Figure Lengend Snippet: Multiplex bead-based EV flow cytometry assay for surface markers. EVs enriched from UHPi and IVIg by SEC and dUC. Surface marker profiles were measured using Miltenyi MACSPlex EV kit IO with MESF-calibrated flow cytometry. (A) Heatmap and hierarchical clustering of MACSPlex markers in UHPi EVs, IVIg EVs, and bead/antibody controls (left). Median APC intensity (MESF, background-subtracted) of mixed tetraspanin antibodies (CD9/CD81/CD63) on EVs captured by 39 marker beads (middle/right). (B) Enlarged values for the tetraspanins (CD9, CD63, and CD81), HLA markers (HLA-ABC and HLA-DR, DP,DQ), platelet markers (CD42a, CD41b, and CD62p), and stemness markers (CD29, ROR1, CD24, CD326, CD133/1). dUC, differential ultracentrifugation; EV, extracellular vesicles; Human Leukocyte Antigens; HLA; IVIg, intravenous immunoglobulin; MESF, Molecular Equivalents of Soluble Fluorophore; SEC, size-exclusion chromatography; UHPi, individual unprocessed human plasma.

Article Snippet: EVs isolated from UHPi, UHPp, and IVIg were subjected to immune profiling for 39 known EV surface proteins by flow cytometry using the human MACSPlex EV kit IO (Miltenyi Biotec, Auburn, CA) and detected by flow cytometry (Cytek Aurora, Cytek Biosciences) with fluorescence calibration using Molecular Equivalents of Soluble Fluorophore (MESF) standards (Quantum APC MESF Beads, Bangs Laboratories) as previously described ( ; ).

Techniques: Multiplex Assay, Flow Cytometry, Marker, Size-exclusion Chromatography, Clinical Proteomics

Journal:

Article Title: EBP2 Is a Member of the Yeast RRB Regulon, a Transcriptionally Coregulated Set of Genes That Are Required for Ribosome and rRNA Biosynthesis

doi: 10.1128/MCB.21.24.8638-8650.2001

Figure Lengend Snippet: Gene expression levels can be assessed by a relative-quantitative RT-PCR technique. (A) RT-PCRs with primers specific to the HHT2 gene were performed on a total RNA sample derived from a logarithmically growing culture of strain yMM177. The products were labeled with [α-32P]dATP, separated by gel electrophoresis, and quantified by phosphorimaging analysis. In order to determine the linear range of the assay, levels of [α-32P]dATP incorporation were measured for up to 26 amplification cycles. (B) RT-PCR assays were performed simultaneously with primers sets specific to both the HHT2 and 18S rRNA sequences. Increasing amounts of nonamplifying competimer 18S primers were added to dampen the 18S amplification reactions down to a level comparable to the HHT2 reaction.

Article Snippet: PCRs were then performed as described by Ambion's Quantu mRNA Universal 18S rRNA Internal Standards Kit.

Techniques: Expressing, Quantitative RT-PCR, Derivative Assay, Labeling, Nucleic Acid Electrophoresis, Amplification, Reverse Transcription Polymerase Chain Reaction

Journal:

Article Title: EBP2 Is a Member of the Yeast RRB Regulon, a Transcriptionally Coregulated Set of Genes That Are Required for Ribosome and rRNA Biosynthesis

doi: 10.1128/MCB.21.24.8638-8650.2001

Figure Lengend Snippet: Motifs 1 and 2 are important for the regulated expression of the EBP2 gene after release from alpha factor arrest. Yeast strains containing either the EBP2ΔN62, the p-M1 EBP2ΔN62, or the p-M2 EBP2ΔN62 plasmid-borne alleles were arrested with alpha factor and released into the cell cycle (A) or else subjected to heat shock (B). RNA samples were prepared for RT-PCR analysis and the amplification products derived from the 18S rRNA, the integrated EBP2, and the EBP2ΔN62 alleles were separated and quantitated. Each expression time point value represents the mean expression level derived from at least two experiments.

Article Snippet: PCRs were then performed as described by Ambion's Quantu mRNA Universal 18S rRNA Internal Standards Kit.

Techniques: Expressing, Plasmid Preparation, Reverse Transcription Polymerase Chain Reaction, Amplification, Derivative Assay