651 cd45ro viogreen  (Miltenyi Biotec)

Journal: Science Advances

Article Title: Human precursor T follicular regulatory cells are primed for differentiation into mature Tfr and disrupted during severe infections

doi: 10.1126/sciadv.adv6939

Figure Lengend Snippet: ( A ) Schematic diagram outlining the study design for TPC. Created in BioRender. Wing, J. (2025) https://BioRender.com/qx390a1 . TGF-β, transforming growth factor–β. ( B ) Flow cytometry analysis showing the levels of BCL6 and CXCR5 expression in T regs from different groups. Statistics: Repeated measures one-way ANOVA with a Holm-Sidak posttest, n = 6 (FMO only, n = 3). ( C ) PCA plot visualizing the gene expression profiles of TPC- and IL-2–treated T regs and Tfr cells. ( D ) Volcano plot depicting DEGs between nT reg -TPC (blue) and preTfr-TPC (red) populations. Statistics: edgeR corrected for donor differences, n = 3 per condition. ( E ) mRNA expression level of selected genes. Statistical significance (adjusted P values from edgeR) is indicated with * P < 0.05, ** P < 0.01, and *** P < 0.001. ( F ) Boxplot showing protein expression level of ILRA determined by ELISA of T reg subset SNs. Statistics: Two-tailed paired t test, n = 6. ( G ) Boxplots of plasmablast suppression assay in total PBMCs treated with TPC culture SNs. Statistics: edgeR corrected for donor differences, n = 3 to 4 per condition (unstim, n = 2 but not used for statistical comparison). Hinges correspond to the first and third quartiles, center line is median, and whiskers correspond to the 1.5 times interquartile range. FMO, flourescence minus one; unstim, unstimulated.

Article Snippet: For in vitro cell expansion, 4 × 10 3 CD45RA + CD45RO − CXCR5 − nT reg or 3 × 10 3 CD45RA + CD45RO − CXCR5 + preTfr cells were activated by human T reg expander beads (Miltenyi Biotec) and cytokine cocktails of IL-2 (500 IU/ml; Shionogi), activin A (100 ng/ml; PeproTech), and IL-12 (1 ng/ml; PeproTech).

Techniques: Flow Cytometry, Expressing, Gene Expression, Enzyme-linked Immunosorbent Assay, Two Tailed Test, Suppression Assay, Comparison

Journal: Science Advances

Article Title: Human precursor T follicular regulatory cells are primed for differentiation into mature Tfr and disrupted during severe infections

doi: 10.1126/sciadv.adv6939

Figure Lengend Snippet: ( A ) Schematic diagram outlining the study design for TPC. Created in BioRender. Wing, J. (2025) https://BioRender.com/qx390a1 . TGF-β, transforming growth factor–β. ( B ) Flow cytometry analysis showing the levels of BCL6 and CXCR5 expression in T regs from different groups. Statistics: Repeated measures one-way ANOVA with a Holm-Sidak posttest, n = 6 (FMO only, n = 3). ( C ) PCA plot visualizing the gene expression profiles of TPC- and IL-2–treated T regs and Tfr cells. ( D ) Volcano plot depicting DEGs between nT reg -TPC (blue) and preTfr-TPC (red) populations. Statistics: edgeR corrected for donor differences, n = 3 per condition. ( E ) mRNA expression level of selected genes. Statistical significance (adjusted P values from edgeR) is indicated with * P < 0.05, ** P < 0.01, and *** P < 0.001. ( F ) Boxplot showing protein expression level of ILRA determined by ELISA of T reg subset SNs. Statistics: Two-tailed paired t test, n = 6. ( G ) Boxplots of plasmablast suppression assay in total PBMCs treated with TPC culture SNs. Statistics: edgeR corrected for donor differences, n = 3 to 4 per condition (unstim, n = 2 but not used for statistical comparison). Hinges correspond to the first and third quartiles, center line is median, and whiskers correspond to the 1.5 times interquartile range. FMO, flourescence minus one; unstim, unstimulated.

Article Snippet: For in vitro cell expansion, 4 × 10 3 CD45RA + CD45RO − CXCR5 − nT reg or 3 × 10 3 CD45RA + CD45RO − CXCR5 + preTfr cells were activated by human T reg expander beads (Miltenyi Biotec) and cytokine cocktails of IL-2 (500 IU/ml; Shionogi), activin A (100 ng/ml; PeproTech), and IL-12 (1 ng/ml; PeproTech).

Techniques: Flow Cytometry, Expressing, Gene Expression, Enzyme-linked Immunosorbent Assay, Two Tailed Test, Suppression Assay, Comparison