Journal: Channels (Austin, Tex.)

Article Title: Immunomagnetic separation is a suitable method for electrophysiology and ion channel pharmacology studies on T cells.

doi: 10.1080/19336950.2020.1859753

Figure Lengend Snippet: Figure 2. Comparison of viability and purity after the different separation techniques. Viability was determined with 7-AAD staining while purity was expressed as a percentage of CD4+ cells in the living cell population (see Methods). Measurements were made via flow cytometry, subsequently after the separation (values are plotted as mean±SD for the number of independent experiments indicated in the bar). No significant differences were found among the groups for either viability or purity (one-way analysis of variance, p > 0.05). REAlease®: measured after the complete removal of bead-antibody and REAlease® complex, in label-free configuration (see Figure 1), Negative selection: immunomagnetic negative selection, Sorted: positive selection by flow cytometry.

Article Snippet: Immunomagnetic negative selection-based separation (CD4+ T Cell Isolation Kit Cat. No.: 130–096-533 Miltenyi Biotec B.V. & Co. KG, Bergisch Gladbach, Germany) was prepared to isolate cells without labeling of the CD4 molecule, and these cells served as the “Control” configuration (Figure 1).

Techniques: Comparison, Staining, Flow Cytometry, Selection

Journal: Nature Communications

Article Title: Tamoxifen for the treatment of myeloproliferative neoplasms: A Phase II clinical trial and exploratory analysis

doi: 10.1038/s41467-023-43175-5

Figure Lengend Snippet: a , b Mutant allele burden change (%) in ( a ) responders achieving after 24w tamoxifen treatment allele burden reductions of ≥50% ( n = 3, green) or ≥25%, <50% ( n = 5, orange), and ( b ) in non-responders ( n = 28, red) before treatment, and 12w or 24w after tamoxifen administration. c - d HSPCs measured as colony-forming units in culture (CFU-Cs) from study patients’ peripheral blood mononuclear cells treated with 4OH-TAM (10 mM) or vehicle for 24 h. Reduced ( c ) or unchanged ( d ) HSPC numbers upon ex vivo 4OH-TAM treatment of baseline samples are consistent with the allele burden reductions observed in the same patients after 24w tamoxifen treatment ( c , green ≥ 50%, n = 2; orange≥25%, n = 1; d , red < 25%, n = 5). * p < 0.05, two-tailed unpaired t test. e CFU-C genotyping shows a reduced balance of JAK2 V617F+ colonies, compared with WT colonies, by 4OH-TAM treatment in responders’, but not in non-responders’ samples (allele burden reductions after 24w are marked with green ≥ 50%, n = 1; orange ≥ 25%, n = 2; red < 25%, n = 3). ** p < 0.01, two-tailed paired t test. Data are mean ± SEM. f Heatmap reveals the disparity of gene expression between responders and non-responders at baseline. g , h Integrated pathway enrichment map using gene-sets enriched in responders ( g ) and non-responders ( h ). i , l Gene set enrichment analysis (GSEA) shows a higher activation of HSPCs in responders, with increased expression of ( i ) STAT3-, ( j ) STAT5-, ( k ) inflammation- and ( l ) apoptosis-related genes. NES, normalized enrichment score. FWER, family-wise error rate.

Article Snippet: After ficoll centrifugation of blood, mononuclear cells were subjected to red blood cell lysis and CD34+ HSPCs were immunomagnetically isolated using the Ultrapure CD34 microbead kit (Miltenyi Biotec, Cat. No. 130-100-453) suing AUTOMACS (Miltenyi Biotec).

Techniques: Mutagenesis, Ex Vivo, Two Tailed Test, Gene Expression, Activation Assay, Expressing

Journal: Oncology letters

Article Title: Anticancer effects of curcumin on nude mice bearing lung cancer A549 cell subsets SP and NSP cells.

doi: 10.3892/ol.2018.9488

Figure Lengend Snippet: Figure 1. Light microscopy of A549 cells and SP cells (stained with CD133+) following A549 sorting. (A) Normal cultured A549 cells (x100 magnifica tion). (B) NSP cells were negatively stained by CD133, indicated by a green arrow. SP cells were positively stained by CD133, indicated by an orange arrow (x400 magnification). (C) The sorted SP cells (x100 magnification). (D) SP cells were positively stained by CD133, characterized by brown staining and indicated by an orange arrow (x400 magnification). SP, side population; NSP, non‑SP.

Article Snippet: Briefly, A549 cells were incubated with CD133 immunomagnetic beads (cat. no. 130-100-857; Miltenyi Biotec, Inc., Cambridge, MA, USA) for 30 min at 4 ̊C.

Techniques: Light Microscopy, Staining, Cell Culture