Fig. 6

DNA damage by drug molecules induces IFN-β via hnRNPA2B1-cGAS. a IL-4 conditioned BMDMs were treated with different doses of MTX/Dox or MTX-MPs/Dox-MPs for a different time, and γH2A.X expression was determined by western blot. b IL-4 conditioned BMDMs were treated with T-MPs at different times, and γH2A.X expression was determined by western blot. c IL-4 conditioned BMDMs were treated with T-MPs, MTX or MTX-MPs for 24 h and the cell cycle was detected by flow cytometry. d, e IL-4 conditioned BMDMs were treated with MTX/Dox or MTX-MPs/Dox-MPs for 24 h, p204, hnRNPA2B1 and cGAS expression and location were determined by real-time PCR and western blot. f The same as (d), except that the nuclear or cytoplasmic p204 and Cgas mRNAs were detected by real-time PCR. g, h BMDMs were transfected with hnRNPA2B1 siRNAs for 12 h and stimulated with IL-4 for 12 h, and treated with MTX or MTX-MPs for 24 h, cGAS and hnRNPA2B1 expression was determined by real-time PCR and western blot (g). IFN-β expression was determined by real-time PCR and ELISA (h). i, j the same as (g, h), except that IL-4 conditioned BMDMs were transfected with Cgas siRNAs. k IL-4 conditioned BMDMs were treated with MTX or MTX-MPs for 24 h, followed by western blot analysis of STING, TBK1, and IRF3. l BMDMs were transfected with hnRNPA2B1 or Cgas siRNAs for 12 h and stimulated with IL-4 for 12 h, and treated with MTX or MTX-MPs for 24 h, followed by western blot analysis of TBK1 and IRF3. Unless otherwise specified, n = 3 biologically independent experiments were performed. Data are presented as mean ± SEM. P values were calculated using one-way ANOVA. ***P < 0.001, ****P < 0.0001