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. Author manuscript; available in PMC: 2020 Apr 24.
Published in final edited form as: Cell Rep. 2019 Oct 29;29(5):1203–1220.e7. doi: 10.1016/j.celrep.2019.09.064

Figure 6. Batf Is Required for Chromatin Looping at the Il21 Locus in Activated CD4+ T Cells.

Figure 6.

(A–C) Chromosome conformation capture (3C) assay performed on 24-h and 96-h WT Th0 and Th0+IL-6 cells (A–C) and 96-h Batf KO cell (B and C) shows relative crosslinking frequencies between selected anchor fragments for the Il21 locus (promoter, +28 and −41; A and B) and Il17a/f locus (Il17a conserved non-coding sequence +11; C) and EcoRI fragments containing the indicated regions. Crosslinking frequencies were corrected for differences in ligation and PCR efficiency using reference DNA and normalized to control interaction frequencies with primer pairs within Gapdh locus.

(D–F) Naive WT and Batf KO cells were activated under Th0+IL-6 conditions. On day 2, cells were transduced with control retrovirus vector (Thy1.1) or retroviral vector expressing Ets1-Thy1.1. Thy1.1-positive cells sorted on day 4 were used to assess gene expression by RT-PCR (D), Ctcf binding by ChIP-qPCR (E), and chromatin looping between Il21 promoter and indicated regions by 3C assay (F).

Data are means ± SEM of three to five independent experiments with one individual mouse per experiment. *p < 0.05 (Student’s t test or one-way ANOVA followed by Tukey’s test).