Journal: bioRxiv

Article Title: Clinical-grade cryopreservation unlocks transplant-ready human pancreatic and stem cell–derived islets for diabetes therapy

doi: 10.64898/2026.04.25.720819

Figure Lengend Snippet: ( A, B ) Bulk RNA-seq volcano plots comparing fresh versus VR native islets ( A ) and SC-islets ( B ). ( C ) Heatmap of differentially expressed genes in VR versus control SC-islets. ( D, E ) Pathway analyses highlighting enrichment of apoptosis, cytokine signaling, and stress responses in SC-islets (PANTHER categories; box-and-whisker plots). ( F ) Confocal images of Annexin V staining in VR islets and SC-islets, indicating apoptosis. Scale bar = 75 µm. ( G ) qPCR analysis of apoptosis-related gene expression in VR-treated SC-islets (mean log₂ fold change [log₂FC], n = 3–4 per group; Student’s t -test, * = p < 0.05). Abbreviations: FDR, false discovery rate; HSR, heat shock response; ISR, integrated stress response; OSR, oxidative stress response; SC, stem cell; UPR, unfolded protein response; VR, vitrified and rewarmed.

Article Snippet: Then, 5× Annexin V binding buffer (Cat. #99902, Biotium) was diluted in deionized water to prepare 1× binding buffer.

Techniques: RNA Sequencing, Control, Whisker Assay, Staining, Gene Expression

Journal: Journal of Virology

Article Title: Foot-and-Mouth Disease Virus Exhibits an Altered Tropism in the Presence of Specific Immunoglobulins, Enabling Productive Infection and Killing of Dendritic Cells ▿

doi: 10.1128/JVI.02180-10

Figure Lengend Snippet: FMDV infection of moDC is closely associated with cell death and reduced ability to stimulate FMDV-specific T cell proliferation. (A) MoDC were exposed to medium, O1K-Cad2, or O1K-Cad2 IC at an MOI of 10 for 6 h. Phase-contrast microscopy captured the high levels of dying cells, including some exhibiting classical apoptotic morphology (high power; right). (B) MoDC were exposed to medium, O1K-Cad2, or O1K-Cad2 IC at an MOI of 10 for 1 to 8 h. Samples of each culture were labeled with antibodies against FMDV NS protein 3A or with annexin V conjugated to APC, and PI. Death was defined as staining with annexin V, PI, or both. The data presented are relative to medium-only values; R = 0.929 (Pearson's correlation coefficient) for the linear relationship between the proportion of cells positive for FMDV NS proteins and the proportion dying. (C) Purified CD4+ T cells from an FMDV-vaccinated animal were added to autologous moDC at a ratio of 1 DC to 100 T cells, with either medium, a sample of the antigen used for vaccination, purified O1K-Cad2, or purified O1K-Cad2 in IC at an MOI of 5. After 5 days, [3H]thymidine was added to each well and incubated overnight before harvesting and liquid scintillation counting of the cells for incorporated radioactivity. Pokeweed mitogen (PWM) was used as a positive control, and in all cases, counts exceeded 350,000 cpm (not shown). A representative data set is presented as cpm ± SD. *, P = 0.02; Student's t test. All 3 replicates using cells from 2 different animals were significant at this level or above.

Article Snippet: To detect dying cells in moDC cultures, 5 × 10 5 cells were pelleted by centrifugation and resuspended in annexin V binding buffer (MBL, Nagoya, Japan).

Techniques: Infection, Microscopy, Labeling, Staining, Purification, Incubation, Radioactivity, Positive Control

Journal: Nature Chemical Biology

Article Title: Menin-MLL Inhibitors Reverse Oncogenic Activity of MLL Fusion Proteins in Leukemia

doi: 10.1038/nchembio.773

Figure Lengend Snippet: (a) MTT cell viability assay in the MLL leukemia cells KOPN-8 and MV4;11 induced by MI-2, MI-3 and MI-nc after 72h treatment. Non-MLL leukemia cell line ME-1 is shown for comparison. Data represent mean values for four samples ± s.d. Experiment was performed three times. (b) Apoptosis and cell death induced by MI-2, MI-3 and MI-nc in MV4;11 cells as detected by flow cytometry using AnnexinV/propidium iodide (PI) staining. Data represent mean values for triplicates ± s.d. (c) Selected histograms from cell cycle analysis performed by FACS after PI staining in MV4;11 cells treated with DMSO, MI-2 or MI-nc. (d) Dose-dependent effect of MI-2 on cell cycle progression measured by FACS in MV4;11 cells after PI staining, with MI-nc as a negative control. Data represent mean values for triplicates ± s.d. (e) Wright-Giemsa stained cytospins on THP-1 and MV4;11 cells after 10 days of treatment with DMSO, MI-2 or MI-nc. (f) Detection of CD11b expression in THP-1 cells assessed by flow cytometry after 6 days of treatment with DMSO, MI-2 or MI-nc. Data represent mean values for triplicates ± s.d. (g) Expression of the HOXA9 and MEIS1 genes normalized to 18S rRNA determined by qRT-PCR in THP-1 cells treated for 6 days with MI-2 and MI-3. Data represent mean values for duplicates ± s.d. Experiment was performed three times.

Article Snippet: After incubation, 1.5×10 5 cells were harvested and resuspended in 100 μl 1× Annexin V binding buffer from the Annexin V-FITC Apoptosis kit (BD Biosciences Pharmingen), incubated with 4 μl of AnnexinV-FITC and 6 μl of Propidium iodide (Sigma-Aldrich) at room temperature in the dark for 10 minutes and analyzed by flow cytometry on a LSR II instrument.

Techniques: Viability Assay, Flow Cytometry, Staining, Cell Cycle Assay, Negative Control, Expressing, Quantitative RT-PCR

Journal: Frontiers in Physiology

Article Title: Protonation of Piezo1 Impairs Cell-Matrix Interactions of Pancreatic Stellate Cells

doi: 10.3389/fphys.2020.00089

Figure Lengend Snippet: Yoda1 impairs PSC viability in spheroids in acidic conditions. (A) Representative Mn 2+ quench traces of PSCs upon acidification with 30 mM sodium propionate (PA) in pH 6.6 or pH 7.4 Ringer's solution. Scatter plots show statistical evaluation in each condition ( n = 48, n = 57, n = 45, n = 46, respectively, N = 5). (B) Microbead traction toward the spheroid in the presence of vehicle (0.1% DMSO, Ctrl) or 20 μM Yoda1 in pH 6.6 or pH 7.4 buffered medium supplemented with 50 ng/ml PDGF ( n = 48, n = 57, n = 45, n = 46, respectively, N = 5). (C) Total area of dissociated acellular space within a spheroid section compared to whole cross-sectional spheroid area ( n = 58, n = 42, n = 35, n = 52 sections, respectively, of spheroids from N = 3 mice). (D) Representative spheroid sections stained with hematoxylin and eosin. (E,F) Scatter plots indicate viability of PSC spheroids in conditions detailed above using MTT reduction assay and Annexin V staining, respectively ( n = 6, N = 3). Data points were normalized to pH 7.4 buffered medium supplemented with 50 ng/ml PDGF and vehicle (0.1% DMSO). * p < 0.05.

Article Snippet: Briefly, PSC spheroids were extracted from the collagen gels manually, washed with PBS and then inserted into tubes containing 0.25% Trypsin-EDTA solution (Sigma-Aldrich, Merck KGaA, Darmstadt, Germany) and shaken at 250 rpm at 37°C for 10 min. After centrifugation at 200 g for 5 min, cells were resuspended in Annexin V binding buffer (Mybiosource, San Diego, CA, USA) and stained with Annexin V conjugated with Alexa 555 (A35108, Invitrogen, Carlsbad, CA, USA, 1:20) at RT for 15 min. Lastly, cells were washed with Annexin V binding buffer and visualized using a fluorescence microscope at 10× magnification in the red fluorescence channel and DIC (Zeiss Axio Observer, Zeiss, Oberkochen, Germany).

Techniques: Staining, MTT Reduction Assay