Supplemental figure A FA-D2 +FANCD2 FA-D2 +FANCD2(K561R) No HU 0.1mM HU S phase: 43%S phase: 34.7% S phase: 30.8% S phase: 52.7%S phase: 25.4%S phase:

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Supplemental figure A FA-D2 +FANCD2 FA-D2 +FANCD2(K561R) No HU 0.1mM HU S phase: 43%S phase: 34.7% S phase: 30.8% S phase: 52.7%S phase: 25.4%S phase: 21.9%

Supplemental figure B Input IgG FlagAb IP Rad18 Rad51 FACND2 Blot: 1 2 3

Supplemental figure C - HU Whole cell Blot: CRad51siRNA - HU Chromatin Blot: CRad51siRNA Rad51 Ku86 USP1 Rad51 Ku86 USP

Supplemental figure D Blot: - MMC HU - MMC HU - MMC HU Chromatin Rad51-7Rad51-2siRNAC Blot: - MMC HU - MMC HU - MMC HU Chromatin FANCD2-18FANCD2-2siRNAC PCNA Ku86 Ub-PCNA Ku86 PCNA Ub-PCNA FANCD2 Blot: - MMC HU - MMC HU - MMC HU Whole cell Rad51-7Rad51-2siRNAC Blot: - MMC HU - MMC HU - MMC HU Whole cell FANCD2-18FANCD2-2siRNAC Rad51 Ku86 FANCD2 PCNA Rad51 Ku86 FANCD2 PCNA

Supplemental figure D (continued) Blot: - MMC HU - MMC HU - MMC HU - MMC HU - MMC HU Soluble FANCD2-18 FANCD2-2 siRNAC Rad51-7Rad51-2 PCNA Ku86 FANCD2 PCNA Long exposure

Supplemental figure E PCNA Ku86 - HU DMSOB02 DMSO B02 Whole cell Chromatin Blot: - HU PCNA Ku86 Blot: Ub-PCNA

Supplemental figure F PCNA Ku86 - HU BRCA2-/-BRCA2+/+ BRCA2-/- BRCA2+/+ Whole cell Chromatin Blot: - HU PCNA Ku86 Blot: Ub-PCNA

Supplemental figure G Blot: - MMC HU - MMC HU - MMC HU - MMC HU - MMC HU Chromatin FANCD2-18 FANCD2-2 siRNAC Rad51-7Rad51-2 Blot: - MMC HU - MMC HU - MMC HU - MMC HU - MMC HU Whole cell FANCD2-18 FANCD2-2 siRNAC Rad51-7Rad51-2 PolH Ku86 FANCD2 Rad51 Ku86 FANCD2 PolH

Supplemental figure H Ku86 Blot: Rad51 siRNAC 7 2 Rad51