Figure 4 : Trex1 knockdown restores cytosolic dsDNA accumulation and induction of ISGs in TSA cells treated with 20 Gy.

From: DNA exonuclease Trex1 regulates radiotherapy-induced tumour immunogenicity

Figure 4

(a–c) TSAshNS and TSAshTrex1 cells were treated with doxycycline to induce Trex1 knockdown and tRFP expression, then exposed to a radiation dose of 8 Gy or 20 Gy. (a) After 24 h cells were fixed, permeabilized and stained with an antibody specific for dsDNA, followed by detection with Alexa fluor 488-conjugated secondary antibody. Representative micrographs show DAPI-stained nuclei (blue), cytoplasmic dsDNA (green), cytoplasmic tRFP (red), and the three channels combined. Magnification: × 400; duplicate; n=3 per group. White bars, 10 μm. (b,c) After 24 h cells were harvested for measurement of cytoplasmic dsDNA (b) or Trex1, Ifnb1 and Mx1 expression by qRT-PCR (c). Duplicate; *P<0.05; **P<0.005; ***P<0.0005: t-test; n=3. All data are mean±s.e.m.