Journal: bioRxiv

Article Title: Efficient Generation of Functional TCRαβ + Cytotoxic T Cells from hiPSCs via Small-Molecule Modulation

doi: 10.64898/2026.03.31.715684

Figure Lengend Snippet: A: Schematic overview of the screening approach. hiPSC-derived CD34 + hemogenic endothelial cells were differentiated towards ProT cells for 14 days in a stroma-free protocol . At iT day 14 cells were harvested and replated into a 384 well plate and exposed to a small molecule library. After 7 days cells were stained with anti CD56, anti CD4, anti CD7 with 7-AAD and Calcein Violet viability dyes. Scale bar indicates 20 µm. B: Primary screening results. 5 hits were identified when a Z-score threshold of 1 was applied. C: Hit validation, bar graphs depicting mean percentage of CD4 + cells out of CD45 + population on iT day 28 in hiPSC line EZH1 mut/wt (screening line), BCH1157 and BCH1566. Error bars depict SEM of replicate wells, one way ANOVA comparison against DMSO. E: Exemplary flow cytometry plots on iT day 28 demonstrating an increased proportion of CD4 + CD8 + DP cells after GNF351 treatment in BCH1157 hiPSC-derived ProT cells. F: Mean values plotted from two independent experiments with either continuous drug treatment (solid circles) or one time drug exposure (open circles). SEM of technical replicates is indicated by blue shading. Two-way ANOVA with Dunnett’s multiple comparison test against respective untreated cells.

Article Snippet: The cells were then filtered through a filter-top polystyrene flow cytometry tube (Fisher Scientific, #08-771-23), washed with flow cytometry buffer (PBS + 2% FBS + antibiotics), and centrifuged at 300xg for 5 minutes followed by antibody staining and flow cytometric analysis for expression of CD45 and CD19.

Techniques: Derivative Assay, Staining, Biomarker Discovery, Comparison, Flow Cytometry

Journal: International Journal of Molecular Sciences

Article Title: Functional Characterization of VS-186B, a Novel HDAC Inhibitor with Anticancer Activity

doi: 10.3390/ijms262311354

Figure Lengend Snippet: ( a ) Phosphatidylserine externalization was observed after 18 h of exposure to VS-186B, indicating apoptotic cell death. Cells were stained with Annexin V FITC and PI and analyzed through flow cytometry. The x-axis represents negative (untreated), vehicle (DMSO), and positive (H 2 O 2 ) controls, as well as VS-186B at its 24 h CC 50 (3.63 µM) and CC 50 × 2 (7.26 µM). The y-axis dictates the percentage of cells undergoing apoptosis. The summation of both early and late apoptotic subpopulations determines the total percentage of apoptotic cells. Representative flow cytometry plots demonstrate viable cells, early apoptotic cells, late apoptotic cells, and necrotic cells after CC 50 and DMSO treatment at 24 h. ( b ) After 18 h of exposure to VS-186B, there is significant ROS production. Flow cytometry plots indicate the percentage of cells induced by ROS after CC 50 and DMSO treatment. Gray represents the negative and vehicle controls and red represents the positive control and treatments. ( c ) Jurkat cells were treated with VS-186B for 10 h, and mitochondrial membrane depolarization was assessed by JC-1 staining and flow cytometry analysis. Flow plots indicate percentage of cells with depolarized mitochondria after CC 50 and DMSO treatment. ( d ) Treatment of VS-186B for 10 h resulted in caspase-3/7 activation, supporting the activation of intrinsic apoptotic cell death. Flow plots represent active caspase percentage comparing CC 50 and DMSO. Gray represents the negative and vehicle controls and red represents the positive control and treatments. Data represents the mean ± standard deviation (SD) of three independent measurements. A two-tailed Student’s paired t -test was performed by comparing DMSO to VS-186B samples. * p < 0.05, ** p < 0.01, *** p < 0.001. Black arrows point to the cell subpopulations shown in each flow cytometry plot.

Article Snippet: 400 μL of cell suspension was distributed into two flow cytometry tubes (unstained and PI-stained) and 5 μL of Propidium Iodide (PI; MP Biomedicals, Solon, OH, USA) at a final concentration of 5 μg/mL was added to each tube, and samples were analyzed by flow cytometry.

Techniques: Staining, Flow Cytometry, Positive Control, Membrane, Activation Assay, Standard Deviation, Two Tailed Test

Journal: International Journal of Molecular Sciences

Article Title: Functional Characterization of VS-186B, a Novel HDAC Inhibitor with Anticancer Activity

doi: 10.3390/ijms262311354

Figure Lengend Snippet: Cell Cycle Analysis of Jurkat Cells Treated with VS-186B: Jurkat cells were treated for 24 h with VS-186B at CC 10 (0.726 µM), CC 20 (1.45 µM), and CC 30 (2.18 µM). No cell cycle phase arrests were observed at these concentrations; however, Sub-G 0 -G 1 populations indicate DNA fragmentation. PI was used to measure DNA content by flow cytometry. Data represents the mean ± standard deviation (SD) of technical triplicates.

Article Snippet: 400 μL of cell suspension was distributed into two flow cytometry tubes (unstained and PI-stained) and 5 μL of Propidium Iodide (PI; MP Biomedicals, Solon, OH, USA) at a final concentration of 5 μg/mL was added to each tube, and samples were analyzed by flow cytometry.

Techniques: Cell Cycle Assay, Flow Cytometry, Standard Deviation