Fig. 7

Derepression of TRIM28 leads to increased synthesis of immunostimulatory cytokines. (A) Bone marrow-derived monocyte (BM) differentiated into dendritic cells (DC) upon culture with GM-CSF for 5 d. Conditioned medium from necroptotic cells (CM) or control cells was added for further activation. (B) DCs were treated with CM for 16 h, and CD86, CD40, and CD80 mRNA levels were analyzed by qPCR. (C) DCs were treated with CM for 16 h, and CD86 expression levels were analyzed by FACS. (D) DCs were treated with CM from L929 cells, which were treated with TZ for 120 min in the absence or presence of Nec-1 (CM), for 16 h. FACS analysis was performed. (E) DCs from RIPK3 WT or RIPK3 KO mice were treated with CM. CD86 expression was analyzed by FACS. (F and G) DCs were treated with CM from L929 cells stably expressing TRIM28 shRNA or the non-silencing control for 16 h. CD86 and MHC II expression was analyzed by FACS (F), and CD86, CD40 and CD80 mRNA levels were analyzed by qPCR (G). (H) The heatmap table shows the association between TRIM28 or RIPK3 and tumor-infiltrating level of multiple types of CD8⁺ T cell or DC, and that is estimated by six algorithms across cancer types in the TCGA database. Red indicates a statistically significant positive association, and blue indicates a statistically significant negative association. (I) Schematic of proposed RIPK3/TRIM28 activation-mediated immunostimulatory cytokine production