Journal: Science Advances
Article Title: TDRD3, a Tudor domain-containing protein, regulates Klf2 -dependent T reg differentiation and function to modulate immune tolerance
doi: 10.1126/sciadv.aea3960
Figure Lengend Snippet: ( A and B ) Immunoblot analysis of TDRD3 in CD4 + YFP + T regs (A) or CD4 + cells (B) isolated from the spleen of indicated mice. Right: qPCR analysis of Tdrd3 mRNA ( n ≥ 3). ( C ) Representative flow cytometric analysis (left two panels), percentage (third panel) and number (right panel) of T reg (Foxp3 + ) cells in thymocytes from indicated mice ( n ≥ 6). ( D and E ) Representative flow cytometric analysis (left two panels) and percentage (third panel) of Foxp3 + iT regs among CD4 + T cells from Foxp3 YFP-Cre or Tdrd3 fl/fl /Foxp3 YFP-Cre mice (D) or from Tdrd3 fl/fl and Tdrd3 fl/fl /CD4 Cre mice (E), polarized in the presence of TGF-β (5 ng/ml) for 48 hours ( n ≥ 6 per treatment cohort). Foxp3 mRNA levels in differentiated iT regs were detected by qPCR (right). ( F ) Representative flow cytometric analysis (two left panels) and percentage (right panel) of live cells among Foxp3 + iT regs differentiated from naïve CD4 + T cells of indicated mice ( n ≥ 6). ( G ) Representative flow cytometric analysis (two left panels) and percentage (right panel) of the proliferating Foxp3 YFP-Cre or Tdrd3 fl/fl /Foxp3 YFP-Cre CD4 + T cells in indicated peaks shown on left, labeled with CellTrace Violet (CTV) and polarized under T reg conditions for 48 hours ( n ≥ 6). Boxed area: cell population of interest. Data are from three independent experiments [(A), (B), (C), (E), (F), and (G), right panels; presented as means ±SEM] or are from one representative of three independent experiments [(A), (B), (C), (E), (F), and (G), left panels]. ** P < 0.01, *** P < 0.001, and **** P < 0.0005; ns, not significant (two-tailed Student’s t test).
Article Snippet: Recombinant TGF-β (#130-095-067) was from Miltenyi Biotec.
Techniques: Western Blot, Isolation, Labeling, Two Tailed Test
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![( A and B ) Immunoblot analysis of TDRD3 in CD4 + YFP + T regs (A) or CD4 + cells (B) isolated from the spleen of indicated mice. Right: qPCR analysis of Tdrd3 mRNA ( n ≥ 3). ( C ) Representative flow cytometric analysis (left two panels), percentage (third panel) and number (right panel) of T reg (Foxp3 + ) cells in thymocytes from indicated mice ( n ≥ 6). ( D and E ) Representative flow cytometric analysis (left two panels) and percentage (third panel) of Foxp3 + iT regs among CD4 + T cells from Foxp3 YFP-Cre or Tdrd3 fl/fl /Foxp3 YFP-Cre mice (D) or from Tdrd3 fl/fl and Tdrd3 fl/fl /CD4 Cre mice (E), polarized in the presence <t>of</t> <t>TGF-β</t> (5 ng/ml) for 48 hours ( n ≥ 6 per treatment cohort). Foxp3 mRNA levels in differentiated iT regs were detected by qPCR (right). ( F ) Representative flow cytometric analysis (two left panels) and percentage (right panel) of live cells among Foxp3 + iT regs differentiated from naïve CD4 + T cells of indicated mice ( n ≥ 6). ( G ) Representative flow cytometric analysis (two left panels) and percentage (right panel) of the proliferating Foxp3 YFP-Cre or Tdrd3 fl/fl /Foxp3 YFP-Cre CD4 + T cells in indicated peaks shown on left, labeled with CellTrace Violet (CTV) and polarized under T reg conditions for 48 hours ( n ≥ 6). Boxed area: cell population of interest. Data are from three independent experiments [(A), (B), (C), (E), (F), and (G), right panels; presented as means ±SEM] or are from one representative of three independent experiments [(A), (B), (C), (E), (F), and (G), left panels]. ** P < 0.01, *** P < 0.001, and **** P < 0.0005; ns, not significant (two-tailed Student’s t test).](https://pub-med-central-images-cdn.bioz.com/pub_med_central_ids_ending_with_9583/pmc12829583/pmc12829583__sciadv.aea3960-f1.jpg)