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PTIP·PA1 localization depends on a signaling cascade involves H2AX, MDC1, and RNF8. HeLa cells or HeLa cells stably expressing HA-FLAG-PTIP were transfected with control siRNA or H2AX-, MDC1-, RNF8-, PTIP-, PA1-specific siRNAs. Cells were irradiated (10 Gy), fixed, and immunostained with anti-FLAG, anti-PA1, and pH2AX antibodies. Western blot analysis was carried out to verify the knockdown of H2AX, MDC1, and RNF8 (lower panel).
