Fig. 2: ATR inhibition induces replication stress-associated DNA damage, genomic instability, apoptosis and cell cycle disruption.
From: Therapeutic targeting of ATR in alveolar rhabdomyosarcoma
Western immunoblot of RPA32 phosphorylation at T21 in Rh4 cells treated with ATR inhibitor AZD6738 (750 nM) (a) and BAY 1895344 (20 nM) (b). c Quantification of TUNEL signal in cells treated with AZD6738 for 72 h. (n = 3; from left to right, P = 5.97 × 10−6; 6.51 × 10−4; 0.002; 0.001; 6.88 × 10−6; 9.04 × 10−4; 0.734; 0.980). d Representative photomicrographs of micronucleation in cells. White arrow represents micronuclei. e Fraction of micronucleated cells after treatment with AZD6738 for 72 h. (n = 3, with 50 nuclei counted per replicate; P = 0.007; 0.007; 0.004; 0.007; 0.007; 0.004; 0.206; 0.768). f Fraction of apoptotic cells after treatment with AZD6738 for 72 h. (n = 3; from left to right, P = 4.54 × 10−9; 7.12 × 10−4; 6.12 × 10−6; 2.46 × 10−4; 6.52 × 10−5; 0.313; 0.424; 0.713). g Cell cycle phase distribution of cells after treatment with AZD6738 for 72 h. (n = 3). Western immunoblot of histone 3 phosphorylation at S10 in six FP-RMS cells treated with AZD6738 (h) and BAY 1895344 (i) for 2 h. j Quantification of changes in histone 3 S10 phosphorylation (P = 0.344; 0.016; statistical analysis is sign test). k Fraction of aneuploid cells after treatment with AZD6738 for 72 h. (n = 3; from left to right, P = 2.55 × 10−5; 5.45 × 10−4; 6.56 × 10−5; 0.402; 5.13 × 10−4; 0.012; 0.882; 0.565). All statistical analyses correspond to two-sided student’s t-test except for (j) data presented as mean value ± error bars representing standard deviation.
