Fig. 7: The Zfp335/Ankle2/Baf axis suppresses cGAS/STING-mediated apoptosis of DN4 thymocytes.
From: Loss of Zfp335 triggers cGAS/STING-dependent apoptosis of post-β selection thymocytes
a Schematic diagram of inhibitors (RU.521 or H-151) or agonists (CMA) used to study cGAS/STING-dependent apoptosis of DN4 thymocytes. Representative histograms (b) and quantification (c) of Annexin V binding for WT (n = 5 mice) or Zfp335cKO (n = 4 mice) DN4 thymoctes treated with cGAS (RU.521) or STING (H-151) inhibitors or vehicle control and cultured on OP9-DL1 stromal cells for 3 days. Total thymocyte (d), DN, DP, CD4SP, and CD8SP or DN1-DN4 cell numbers (e, f) or frequencies (g, h) for Zfp335cKO mice treated with H-151 (n = 8 mice) or vehicle (n = 6 mice) in vivo for 7 days. i Quantification of Annexin V binding among DN4 cells from R26LSL-Cas9 TcrdCreERT2 thymocytes transduced with gRNA-expressing retroviruses and cultured for three days on OP9-DL1 cells with 4-hydroxytamoxifen. n = 3 biological replicates pooled from 3 mice each for each dual gRNA transduction. Percent apoptosis induced by small molecule activation of STING among WT thymocyte subsets treated with 25 µg/mL (j) or 250 µg/mL CMA (k), and Zfp335cKO Bcl2-Tg (l), or αCD3-treated Rag2−/− thymocyte subsets (m) treated with 25 µg/mL CMA. Panels j and k were from the same experiments. n = 6 mice (j, k), n = 8 mice (l), or n = 7 mice (m). P-values determined by two-tailed Mann–Whitney U-test (d) or two-way ANOVA with post hoc Tukey’s test (c) or Sidak’s test (e–h, j–m) or one-way ANOVA with post hoc Tukey’s test (i). Data shown are compiled from two (b, c, m), three (i–l), or five (d–h) independent experiments utilizing male and female mice. Plots show mean ± SEM. Source data are provided as a Source Data file.
