Fig. 6. ZFP36 and ZFP36L1 limit the early expression of transcription factors driving effector differentiation.

a CLIP data showing sequencing reads across (a) Nfkb2 and (b) Rel transcripts (a set of top two lanes) with an expanded view of the 3ʹUTR (a set of bottom two lanes). In each set top lane shows ZFP36L1 CLIP data from OT-I CD8 CTLs stimulated for 3 h with N4 peptide; bottom lane shows ZFP36 CLIP data from in vitro activated naive CD4 T cells. TATTTA motifs are identified in orange; (c, d) conservation among vertebrates of the ZFP36/ZFP36L1 binding site determined using MULTIZ alignment tool in UCSC genome browser for (c) Nfkb2 and (d) Rel. Binding as identified by CLIP is highlighted in yellow. Selected species are ordered from top to bottom according the evolutionary proximity of the clades. Red bracket highlights the ARE. For Rel the best conserved binding site is displayed. e Differential protein expression of NF-κB2 and (f) cREL by naive CD8⁺ T cells following activation with plate bound anti-CD3 antibody. Representative flow cytometry histograms (left panel) and geometric mean fluorescence (right panel) are shown. Open histograms represent WT and filled histograms show dKO cells. Statistical significance was determined using a mixed-effects model analysis followed by Sidak’s test for multiple comparisons. Data is compiled from two of three independent experiments, with each dot representing a biological replicate. g Flow cytometry plots showing expression of EOMES and GzmB by naive WT and dKO CD8⁺ T cells activated with plate bound CD3 for 72 h. h Frequency of GzmB and EOMES expressing cells. Statistical significance was tested with a two-tailed unpaired Student’s t-test. Data is compiled from two independent experiments and three biological replicates. In all panels filled circles represent the respective WT control and open shaded circles dKO. Source Data are provided as Source Data file.