Fig. 7: Cul5 regulates fate choice in CD4⁺ T cells.

a Representative flow plots showing the frequencies of FoxP3⁺ and IL-9⁺ cells. Naive CD4⁺ T cells from control and Cul5^fl/flCD4-Cre mice were cultured in iTreg polarizing (TGF-β + IL2 + anti-IFNγ) condition with increasing concentrations of IL-4. Cells were stained for FoxP3 and IL-9 on day 5. b Compiled data of (a) showing the frequencies of FoxP3⁺ (blue) and IL-9⁺ (red) cells. For FoxP3⁺ cells, n = 3 control, n = 4 Cul5^fl/flCD4-Cre, for IL9⁺ cells n = 4 control, n = 5 Cul5^fl/flCD4-Cre biologic replicates of cells examined in three independent experiments. c Compiled data showing the frequencies of FoxP3⁺ cells in 8–10-week-old control and Cul5^fl/flCD4-Cre mice treated with PBS. n = 8 control and Cul5^fl/flCD4-Cre mice examined in two independent experiments. (d) HDM treated mice n = 11 control and Cul5^fl/flCD4-Cre mice examined in three independent experiments. Mixed bone marrow chimeras. 6–8-weeks-old Rag1-/- deficient recipients were sublethally irradiated and then injected with a 1:1 mixture of control (CD45.1⁺) and Cul5^fl/flCD4-Cre (CD45.2⁺) donor bone marrow cells and allowed to reconstitute for 8 weeks. These mice were then treated with a 2-week regimen of house dust mite (HDM), and their lung cells were analyzed by flow cytometry. e Proportion of FoxP3⁺ cells that came from each donor. The percent of control (CD45.1⁺) and Cul5^fl/flCD4-Cre (CD45.2⁺) FoxP3⁺ cells was normalized to the percent of CD4⁺ T cells from the same donor. n = 7 recipient mice were examined in two independent experiments. Two donors of each genotype were used to reconstitute these recipients. Each recipient received cells from one mouse of each genotype. Proportion of cytokine producing cells. To calculate the contribution from each donor CD45.1 “cytokine⁺” cells or CD45.2 “cytokine⁺” cells were normalized to CD19⁺ cells. f Proportion of IL-9⁺ cells from CD45.1 (WT) and CD45.2 (Cul5 deficient), n = 5 recipients were examined in one experiment. g-h Proportion of IL-4⁺ and IL-5⁺ cells, n = 12 recipients were examined in two independent experiments. Data is presented as mean±SEM in panels b–d. In panels e-h data is represented as mean. p-value was calculated by unpaired two tailed t-test used in panels b-d and paired two-tailed t-test in panels e-h. Source data are provided as a Source Data file.