FANCI Regulates Recruitment of the FA Core Complex at Sites of DNA Damage Independently of FANCD2 (original) (raw)
Fig 1
FA core complex forms foci at sites of DNA damage.
(A) U2OS cells were transfected with siControl or siFANCA and treated with mitomycin C (MMC) 60ng/ml for 24h before fixation. Cells were immunostained with the indicated antibodies. (B) Cells were left untreated or treated with 60ng/ml MMC, 2.5 μM cisplatin, 250μM hydroxyurea (HU) or 10μM AZD2281 for 24h, or treated with 10Gy ionizing radiation (IR) 8h before fixation. Then cells were immunostained with anti-FANCA antibody. Representative images are shown. (C) U2OS cells or U2OS expressing 3xFLAG-FANCD2 were treated with MMC 60ng/ml for 24h and immunostained with anti-FANCA, FLAG and γH2AX antibodies. (D) Localized DNA damage induced with a 450nm laser in U2OS cells expressing EGFP-FANCD2. Cells were fixed 30min after irradiation and immunostained with the indicated antibodies. (E) U2OS cells were transfected with the indicated siRNAs, untreated or treated with MMC 60ng/ml for 24h and immunostained with anti-FANCA or anti-FANCG antibodies. Foci/cell were counted using an automated software. Data represent mean values ± SD of three independent experiments. (*) Indicates p < 0.05; (n.s.) indicates no statistical significance (Compared to siControl).