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. Author manuscript; available in PMC: 2020 Jul 15.
Published in final edited form as: Environ Mol Mutagen. 2020 Apr 16;61(6):635–646. doi: 10.1002/em.22372

Fig. 4.

Fig. 4.

THS exposure causes replication stress and activates DSB response machineries. (A) Replication stress response was monitored following exposure of hPFs to the indicated doses of THS for 12 h followed by western analysis with anti-phosphoATR Thr1989 antibody. Tubulin was used as loading control. (B) Induction of DSBs by exposure of hPFs to 2.5% THS for 24 h was analyzed by IF detection of 53BP1 protein, which accumulates in foci at DSBs. SHS (0.4 PE) treatment was used as a positive control. (C) Activation of ATM following exposure of hPFs to 10%, 5% and 2.5% THS for 12 h was analyzed by western with anti-pATM (Ser1981). The same membrane was probed for phosphorylation of histone H2AX using anti-γH2AX antibody (Ser139) as an indicator of DSB formation and for pRPA32 (as in Fig. 3) to monitor replication stress. (D) Micronuclei formation in BEAS-2B cells following exposure to 5% THS for 48 h. Data represent the mean of ±SD for N=2.