Journal: Cancers

Article Title: Euchromatic Histone Lysine Methyltransferase 2 Inhibition Enhances Carfilzomib Sensitivity and Overcomes Drug Resistance in Multiple Myeloma Cell Lines.

doi: 10.3390/cancers15082199

Figure Lengend Snippet: Figure 4. UNC0642/CFZ combination is not toxic for normal cells, and it is effective in the presence of bone marrow milieu. (a) PMBCs were extracted from four healthy donors. Combinatorial treatments with increasing doses of UNC0642 together with 2.5 nM CFZ did not affect the viability of PBMCs compared to myeloma KMS28-BM control cells. Annexin V-PI staining was performed 72 h post- treatment. (b) GFP+ bone marrow stromal cells (HS-5) were seeded at 50,000/mL and subjected to single or double treatment. KMS28-BM cells were plated separately and used as a control. Cell death was monitored with FACS (PI-staining or TMRM-staining) at the indicated time points. (c) The KMS28-BM cell line was cultured on a monolayer of GFP+ HS-5 stromal cells and treated with the indicated doses of UNC0642 and CFZ. Cell viability was estimated by FACS analysis by staining with CD138-APC antibody, and the ratio with GFP+ HS-5 was calculated at the indicated time points. Statistical significance was calculated with the two-way ANOVA test (n = 7). (d) 3D scaffolds were populated with HS-5 cells (GFP+) and RPMI-I8226 cells (tRFP+) and treated with CFZ, UNC0642, or with the combination. RPMI-8226 cells were specifically detected using FACS analysis 6 days after treatment (n = 2 independent replicates) (unpaired t-test CFZ vs. CFZ + UNC0682 p-value = 0.0019). * p <0.05; ** p < 0.01; *** p < 0.001.

Article Snippet: Apoptosis was measured by flow cytometry after staining with tetrametylrodamine methyl ester (TMRM; Molecular Probes, Eugene, OR, USA) or Annexin V-FITC Kit (Catalog No. 130-092-052, Miltenyi Biotec, Bergisch Gladbach, Germany), according to the manufacturer’s instructions.

Techniques: Control, Staining, Cell Culture

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Reduction in the developmental potential of intrathymic T cell progenitors with age.

doi: 10.4049/jimmunol.173.1.245

Figure Lengend Snippet: FIGURE 4. Proliferation and survival of ETP from young and old mice. Thymocytes were freshly isolated from young and old mice and stained for ETP in combination with anti-Ki-67 or annexin V labeling to determine their proliferation or survival status, respectively. A, ETP were labeled with anti-Ki-67 or isotype control Abs following fixation and permeabilization. The upper panel shows representative data for single young and old ani- mals. The lower panel indicates data pooled from the indicated number of different aged mice at each time point. B, The level of apoptosis in ETP was evaluated by annexin V labeling. The upper panel shows representative data for single young and old animals. The lower panel indicates data pooled from the indicated number of different aged mice at each time point. Means and SDs are shown for n 2–6 animals as indicated. , p 0.02; , p 0.002; #, p 0.005; ##, p 0.0005.

Article Snippet: The frequency of apoptotic cells in freshly harvested thymocyte populations was assessed by labeling with annexin V using the Apoptosis Detection kit as described by the manufacturer (R&D Systems, Minneapolis, MN) in combination with Abs to various cell surface determinants.

Techniques: Isolation, Staining, Labeling, Control