Research Paper Volume 13, Issue 12 pp 15833—15874

The lack of functional DNMT2/TRDMT1 gene modulates cancer cell responses during drug-induced senescence

DNMT2/TRDMT1 gene knockout-mediated changes in the levels of NSUN proteins (A, B) in U-251 MG glioblastoma cells treated with DOX or ETOPO. (A) The levels of NSUN1, NSUN2, NSUN3, NSUN4, NSUN5 and NSUN6 are expressed as relative fluorescence units (RFU). Box and whisker plots are shown, n = 3, ***p **p *p a posteriori test), ###p ##p #p a posteriori test). (B) NSUN1, NSUN2, NSUN3, NSUN4, NSUN5 and NSUN6 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.

Figure 8. DNMT2/TRDMT1 gene knockout-mediated changes in the levels of NSUN proteins (A, B) in U-251 MG glioblastoma cells treated with DOX or ETOPO. (A) The levels of NSUN1, NSUN2, NSUN3, NSUN4, NSUN5 and NSUN6 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) NSUN1, NSUN2, NSUN3, NSUN4, NSUN5 and NSUN6 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.