Research Paper Volume 13, Issue 12 pp 15833—15874

Figure 7. DNMT2/TRDMT1 gene knockout-mediated senescence-associated secretory phenotype (SASP) (A, B) in U-251 MG glioblastoma cells treated with DOX or ETOPO. (A) The levels of NF-κB, IL-1β, IL-6 and IL-8 are expressed as relative fluorescence units (RFU). Box and whisker plots are shown, n = 3, ^***p < 0.001, ^**p < 0.01, ^*p < 0.05 compared to CTR (ANOVA and Dunnett’s a posteriori test), ^###p < 0.001, ^##p < 0.01, ^#p < 0.05 compared to C-NIC cells at the same culture conditions (ANOVA and Tukey’s a posteriori test). (B) NF-κB, IL-1β, IL-6 and IL-8 immunostaining (red). Representative microphotographs are shown, objective 20x, nucleus staining (blue), RESPONSE, representative DOX or ETOPO treatment. CTR, control conditions; DOX, doxorubicin treatment; ETOPO, etoposide treatment; AZA, azacytidine treatment; MIX, azacytidine post-treatment; C-NIC, control cells with unmodified levels of DNMT2/TRDMT1 containing control plasmid; D-NIC, cells with DNMT2/TRDMT1 gene knockout containing dedicated DNMT2 double nickase plasmid.