Journal: bioRxiv

Article Title: A CD40-Targeted IL-21 Fusokine Enables Rapid Generation of Human IL-10⁺Granzyme B⁺ Regulatory B Cells

doi: 10.64898/2026.01.12.699061

Figure Lengend Snippet: (A) Principal component analysis (PCA) of RNA-seq data from purified human peripheral blood B cells cultured under four conditions: unstimulated B cells, CpG-stimulated B cells (B-CpG), CD40scFv–IL-21–stimulated B cells (B-ScFv), and B cells stimulated with the combination of CD40scFv–IL-21 and CpG (Bregs). PCA demonstrates clear separation among conditions, indicating distinct global transcriptional states induced by single versus combined stimulation. (B) Venn diagram showing the number of genes upregulated (log₂ fold change > 1) relative to unstimulated B cells in B-CpG, B-ScFv, and Bregs. The overlap highlights both shared and condition-specific transcriptional responses, with Bregs exhibiting a distinct set of upregulated genes not observed with single-stimulus activation. (C) Volcano plot depicting differential gene expression between Bregs and B-CpG cells. The x-axis represents log₂ fold change (Bregs vs B-CpG), and the y-axis represents −log₁₀ adjusted p-value. Genes significantly upregulated in Bregs include regulatory and effector-associated transcripts such as IL10, GZMB, CD274 (PD-L1), PDCD1LG2 (PD-L2) , and EBI3 , whereas CpG-associated inflammatory and chemokine-related genes are less prominent under combined stimulation. (D) Heat map showing expression of selected regulatory, checkpoint, transcriptional, and chemokine-associated genes comparing B-CpG and Bregs across three independent donors (D312, D919, D936). Expression values are displayed as log₂(FPKM + 1) and visualized using a red–blue diverging color scale, with red indicating higher relative expression and blue indicating lower expression. The heat map reveals coordinated upregulation of regulatory and effector-associated genes ( IL10, GZMB, CD274, PDCD1LG2, EBI3, PRDM1, IRF4 ) in Bregs, whereas B-CpG cells show relatively higher expression of chemokines such as CCL3 and CCL4 .

Article Snippet: Chemokine expression was assessed using the Proteome Profiler Human Chemokine Array Kit (R&D Systems, ARY017) according to the manufacturer’s instructions.

Techniques: RNA Sequencing, Purification, Cell Culture, Activation Assay, Gene Expression, Expressing

Journal: Pharmaceuticals

Article Title: Pro-Apoptotic and Anti-EMT Activity of Wild Ginseng Adventitious Root Extract in MDA-MB-231 TNBC Cells: Association with GSK-3β/β-Catenin Signaling

doi: 10.3390/ph19020216

Figure Lengend Snippet: Proteome profiling of oncology-related markers in MDA-MB-231 cells treated with wild ginseng adventitious root extract (WGAR) and cisplatin. Cells were treated with WGAR or cisplatin at the 20% inhibitory concentration (IC20; 3 μg/mL for WGAR, 2 μg/mL for cisplatin) and 50% inhibitory concentration (IC50; 79 μg/mL for WGAR, 9 μg/mL for cisplatin) for 48 h. Cell lysates (200 μg) were analyzed using the Proteome Profiler Human XL Oncology Array (R&D Systems, ARY026). ( a ) Representative array membrane images. ( b ) Quantification of selected oncology-related markers at IC20 concentrations. Proteins are grouped by functional categories as indicated by horizontal lines below the x -axis. ( c ) Quantification of selected oncology-related markers at IC50 concentrations. Proteins are grouped by functional categories as indicated by horizontal lines below the x -axis. ( d ) Independent validation of IL-6 and M-CSF by ELISA. IL-6 and M-CSF levels in culture supernatants were quantified using the Human IL-6 Quantikine ELISA Kit (Cat# D6050, R&D Systems) and Human M-CSF Quantikine ELISA Kit (Cat# DMC00B, R&D Systems), respectively, and expressed as fold change relative to control (set to 1.0). Data are expressed as fold change relative to control (dashed line = 1). Values represent mean ± SD from three independent experiments. Statistical significance was determined by one-way ANOVA followed by Dunnett’s post hoc test: * p < 0.05, ** p < 0.01, *** p < 0.001 compared to the untreated control. CCL2, C-C motif chemokine ligand 2; CXCL8, C-X-C motif chemokine ligand 8; Dkk-1, Dickkopf-1; GM-CSF, granulocyte-macrophage colony-stimulating factor; IL-6, interleukin-6; M-CSF, macrophage colony-stimulating factor; PAI-1, plasminogen activator inhibitor-1; u-PA, urokinase-type plasminogen activator; VEGF, vascular endothelial growth factor; WGAR, wild ginseng adventitious root.

Article Snippet: To analyze the differential expression of multiple cancer-related proteins simultaneously, the Proteome Profiler Human XL Oncology Array Kit (Cat# ARY026, R&D Systems, Minneapolis, MN, USA) was employed according to the manufacturer’s instructions.

Techniques: Concentration Assay, Membrane, Functional Assay, Biomarker Discovery, Enzyme-linked Immunosorbent Assay, Control