u937 human leukaemia  (ATCC)

Journal: Nature Communications

Article Title: A Pseudomonas aeruginosa quorum-sensing metabolite manipulates macrophage ferroptosis through a methylation pathway

doi: 10.1038/s41467-025-65142-y

Figure Lengend Snippet: A Growth curves of PAO1, Δ pqsR , and Δ pqsR + strains. B Lactate dehydrogenase (LDH) assay for cell death in RAW264.7 macrophages infected with PAO1, Δ pqsR , and Δ pqsR + (Δ pqsR mutant expressing pqsR from a rescue construct pMiniCTX1- pqsR ) Bac (bacteria cultures) or Sup (cell-free supernatants). Multiplicity of infection (MOI) = 100. Data represent the means ± SD ( n = 3, two-way ANOVA with Tukey’s multiple comparisons test). C ELISA quantification of PQS concentrations measured in PAO1, Δ pqsR , and Δ pqsR + supernatants. Data represent the means ± SD ( n = 3, one-way ANOVA with Tukey’s multiple comparisons test). D LDH assay for cell death in RAW264.7 macrophages treated with different concentrations of PQS for 24 h, untreated macrophages (Ctrl), and macrophages treated with DMSO as the negative control. Data represent the means ± SD ( n = 3, one-way ANOVA with Tukey’s multiple comparisons test). E CCK-8 assay for cell viability in RAW264.7 macrophages treated with different concentrations of PQS for 24 h; IC 50 (PQS) = 14.98 μg/mL. Data represent the means ± SD ( n = 3). F Microscopic observation of the effect of concentration of IC 50 (PQS) exposure for 24 h on RAW264.7 macrophage morphology. Scale bar = 100 µm. G Transcription electron microscopy (TEM) observation of the effect of 10 μg/mL PQS exposure on RAW264.7 macrophage morphology. Nu: cell nucleus, Au-L: autolysosome. 15/40KK: × 15000/ 40,000 magnifications. Scale bar = 2/0.5 μm. H CCK-8 assay for cell viability in BMDM, THP-1, U937, A549, and BEAS-2B cells treated with 10 μg/mL PQS for 24 h. Data represent the means ± SD ( n = 3). Source data are provided as a Source Data file.

Article Snippet: The RAW264.7 murine macrophages (ATCC ® TIB-71 TM ), THP-1 human monocyte (ATCC ® TIB-202 TM ), U937 human leukaemia (ATCC ® CRL-3253 TM ), A549 human epithelium (ATCC®CCL-185TM), and BEAS-2B human bronchial epithelium (ATCC ® CRL-3588 TM ) cell lines were grown in Dulbecco’s Modified Eagle (DMEM) or Roswell Park Memorial Institute (RPMI)-1640 medium supplemented with 10% foetal bovine serum (FBS) and 1% penicillin/streptomycin (PS) (Gibco, Texas, USA), followed by incubation at 37 °C in a 5% CO 2 incubator.

Techniques: Lactate Dehydrogenase Assay, Infection, Mutagenesis, Expressing, Construct, Bacteria, Enzyme-linked Immunosorbent Assay, Negative Control, CCK-8 Assay, Concentration Assay, Electron Microscopy

Journal: Research

Article Title: Biomimetic Alveolus-on-a-Chip for SARS-CoV-2 Infection Recapitulation

doi: 10.34133/2022/9819154

Figure Lengend Snippet: (a) ROS generation characterization before and after poly(I:C) treatment for ihAEpiC (scale bar, 50 μ m) and HUVEC (scale bar, 100 μ m). Cytokines secreted by (b) ihAEpiC and (c) HUVEC before and after poly(I:C) treatment. (d) Images of U937 cells adhering to HUVEC and number calculation of adhered U937 cells within six random areas in the microchip. Scale bar is 100 μ m.

Article Snippet: U937 human myeloid leukaemia cells were bought from ATCC and cultured in RPMI-1640 medium (Gibco, USA), containing 10% FBS (Gibco, USA) and 1% penicillin-streptomycin (Corning, USA).

Techniques: MicroChIP Assay

Journal: Cell Research

Article Title: Vulnerabilities in mIDH2 AML confer sensitivity to APL-like targeted combination therapy

doi: 10.1038/s41422-019-0162-7

Figure Lengend Snippet: The transcriptional rewiring of mIDH leukaemia evolution is characterized by altered MAPK and ATRA associated pathways. a Gene Set Enrichment Analysis (GSEA) plots for MAPK, TNFα, and ATRA related signatures in third recipient- vs second recipient- derived leukaemia cells. b Heat maps showing the relative abundance of genes specifically enriched in GSEA (a). c Venn diagram showing shared genes between mouse mIDH2 leukaemia and Schenken et al.; 2013 gene sets. d Heat map showing that shared genes shown in c are regulated concordantly in samples derived from second or third recipients with respect to ATRA + TCP (LSD1 inhibitor) gene sets reported by Schenken et al.; 2013. e Histogram showing LSD1 activity measured in nuclear extracts of mIDH2 TF1 (IDH2-R140Q) and relative controls. Data are mean ± SD; of n = 2 samples; unpaired t-test; **p ≤ 0.01. f Western blot assay on TF1 cells overexpressing the mutant R140Q of IDH2 (IDH2R140Q) or respective control cells (CTRL). LSD1 protein levels are slightly reduced in the mutant R140Q of IDH2 (IDH2R140Q). g Quantitative PCR analyses for genes associated with ATRA sensitivity in the context of LSD1 inhibition in human TF1 cells treated for 24 h with ATRA (1 μM) or vehicle (DMSO). ITGAM integrin subunit alpha M, LILRA5 leukocyte immunoglobulin like receptor A5, RARA retinoic acid receptor alpha, PRAM1 PML-RARA regulated adaptor molecule 1. Data are mean ± SD; one -way anova, ***p ≤ 0.001. h Western blot analysis for ATRA targets (Pin1), ATRA receptors (RARα, RXRα), and factors associated with ATRA-related differentiation pathway in hematopoietic cells (c/EBPα, c/EBPε). The samples are protein extracts from independent (n = 2) leukaemia cells isolated from DOX+ (mIDH2 ON) or DOX- (mIDH2 OFF) second or third recipients. Protein samples isolated from Hoxa9/Meis1a leukemic cells (n = 2) were included as control. i–j Western blot analysis for factors associated with ATRA-related pathway in TF1 and U937 cell line, stably overexpressing the mutant isoform R140Q of IDH2 (mIDH2) or the empty vector (CTRL). mIDH2: IDH2R140Q. See also Supplementary Information, Figs. S4 – S6 and Supplementary Information, Table S3

Article Snippet: Cells The human leukaemia U937 and TF1 cell lines were purchased from the American Type Culture Collection (ATCC; https://www.atcc.org ).

Techniques: Derivative Assay, Activity Assay, Western Blot, Mutagenesis, Control, Real-time Polymerase Chain Reaction, Inhibition, Isolation, Stable Transfection, Plasmid Preparation