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