Journal: iScience

Article Title: Elastic properties of leukemic cells linked to maturation stage and integrin activation

doi: 10.1016/j.isci.2025.112150

Figure Lengend Snippet: Measurements of the Young’s modulus of patient samples

Article Snippet: Cryopreserved AML patient samples were thawed and resuspended in newborn calf serum (NCS) supplemented with DNase I (20 Units/mL), 4 mM MgSO 4 and heparin (5 Units/mL) and incubated at 37°C for 15 min. AML MNCs were blocked for 5 minutes with human FcR blocking reagent (Miltenyi Biotec) and stained with antibodies: (CD34-FITC cat#345801, BD Biosciences (2 μl/100 μl); CD38-PE cat#345806, BD Biosciences (3 μl/100 μl); and CD45-PERCP cat#304026, Biolegend (1.5 μl/100 μl)) or with antibodies: (CD34-PE cat#130-120-520, Miltenyi Biotec (2μl /100 μl); CD38-FITC cat#555459, BD Pharmingen (3μl/100μl); and CD45-PE-Cy7 cat#368532, Biolegend (1.5 μl/100 μl)).

Techniques:

Journal: iScience

Article Title: Elastic properties of leukemic cells linked to maturation stage and integrin activation

doi: 10.1016/j.isci.2025.112150

Figure Lengend Snippet: Stiffness measurements of patient AML samples Patient samples were sorted into CD34 + /CD38 + and CD34 + /CD38 - populations and measured using the AFM procedure outlined above. Distributions of the measured Young’s modulus values of cells from (A) patient 1, (B) patient 2, and (C) patient 3. Each point represents the average of 10–14 measurements performed on one cell. Outliers are indicated with the daggers. The horizontal bars show the distribution means and the error bars are the standard error across cells after outlier removal. For all patients, CD34 + /CD38 - cells are significantly stiffer than CD34 + /CD38 + cells. The addition of retronectin has no effect on CD34 + /CD38 + cell stiffness for any of the patients, whereas it has mixed effects on the CD34 + /CD38 - cells. The number of measured cells for each sample, n , is reported in Table 2 . Significance testing was performed using a two sample t test with an alpha of 0.05. The p values are given in Table S4 .

Article Snippet: Cryopreserved AML patient samples were thawed and resuspended in newborn calf serum (NCS) supplemented with DNase I (20 Units/mL), 4 mM MgSO 4 and heparin (5 Units/mL) and incubated at 37°C for 15 min. AML MNCs were blocked for 5 minutes with human FcR blocking reagent (Miltenyi Biotec) and stained with antibodies: (CD34-FITC cat#345801, BD Biosciences (2 μl/100 μl); CD38-PE cat#345806, BD Biosciences (3 μl/100 μl); and CD45-PERCP cat#304026, Biolegend (1.5 μl/100 μl)) or with antibodies: (CD34-PE cat#130-120-520, Miltenyi Biotec (2μl /100 μl); CD38-FITC cat#555459, BD Pharmingen (3μl/100μl); and CD45-PE-Cy7 cat#368532, Biolegend (1.5 μl/100 μl)).

Techniques:

Journal: iScience

Article Title: Elastic properties of leukemic cells linked to maturation stage and integrin activation

doi: 10.1016/j.isci.2025.112150

Figure Lengend Snippet:

Article Snippet: Cryopreserved AML patient samples were thawed and resuspended in newborn calf serum (NCS) supplemented with DNase I (20 Units/mL), 4 mM MgSO 4 and heparin (5 Units/mL) and incubated at 37°C for 15 min. AML MNCs were blocked for 5 minutes with human FcR blocking reagent (Miltenyi Biotec) and stained with antibodies: (CD34-FITC cat#345801, BD Biosciences (2 μl/100 μl); CD38-PE cat#345806, BD Biosciences (3 μl/100 μl); and CD45-PERCP cat#304026, Biolegend (1.5 μl/100 μl)) or with antibodies: (CD34-PE cat#130-120-520, Miltenyi Biotec (2μl /100 μl); CD38-FITC cat#555459, BD Pharmingen (3μl/100μl); and CD45-PE-Cy7 cat#368532, Biolegend (1.5 μl/100 μl)).

Techniques: Recombinant, Sequencing, Software

Journal: Nature Communications

Article Title: CXCL1 mediates obesity-associated adipose stromal cell trafficking and function in the tumour microenvironment

doi: 10.1038/ncomms11674

Figure Lengend Snippet: ( a ) Proliferation of control-shRNA and CXCL1-shRNA RM1 cells growing in 1or 10% FBS assessed by MTT assay. Plotted is relative fluorescence unit (RFE) increase above day 0 value from technical triplicates. ( b ) RM1 cells transduced with an untargeted shRNA (control-sh) or shRNA-targeting CXCL1 (CXCL1-sh) were subcutaneously grafted into non-obese or obese mice ( n =5 per group), and tumour volume was measured weekly. ( c ) Growth of RM1 grafts in mice treated or not treated with reparixin ( n =5 per group) in lean and obese mice. In a – c , graphs show mean±s.e.m.; * P <0.05 (Student's t -test). ( d ) Representative flow cytometric gating to separate tumour CD45− (stromal, vascular and malignant) and CD45+ (haematopoietic) cells. Gating of CD45−CD34+Sca1+ ASCs based on side scatter (SSC) and CXCR1/2 expression reveals increased frequency of CXCR1+ ASCs in control-shRNA, but not in CXCL1-shRNA tumours in obese mice. Gating of CD45− leukocytes based on CD11b and Gr1 expression reveals reduced frequency of CD11b+Gr1+ MDSCs in CXCL1-shRNA tumours in both lean and obese mice. % of total viable cells is shown. Analyses were performed twice with similar results.

Article Snippet: Mouse tumour tissue cell suspensions were analysed as described based on fluorescence or using the following IgG clones: APC-anti-CD34 (RAM34) or PE-anti-CD34 (MEC 14.7), PE-Cy7-anti-CD31 (390) or PE-anti-CD31 (MEC 13.3), APC-Cy7-CD45 (30-F11), CY5.5-anti-Sca-1 (D7), FITC-anti-CXCR1 (orb15459, Biorbyt), PerCP-CXCR2 (FAB2164C, R&D) and the corresponding isotype controls (BD Biosciences).

Techniques: shRNA, MTT Assay, Fluorescence, Transduction, Expressing