Figure EV2. SCAI promotes replication‐coupled ICL repair (related to Fig 3).

• A
  Samples in Fig 3B were replicated in the presence of [α‐³²P]dATP and analyzed by native agarose gel electrophoresis. RI, replication intermediates; OC, open circular; SC, supercoiled.
• B
  SCAI‐C immunoblot analysis of Xenopus egg extracts (NPE) subjected to immunodepletion with SCAI‐C, SCAI‐N, or an IgG control antibody.
• C
  SCAI‐N immunoblot analysis of samples from (B).
• D
  Immunoblot analysis of Xenopus egg extracts subjected to immunodepletion with antibodies targeting the C‐terminal (SCAI‐C) or N‐terminal (SCAI‐N) region of SCAI or an IgG control. Asterisks denote antibody cross‐reactivity. Note that SCAI‐N but not SCAI‐C immunodepletion also co‐depletes CtIP.
• E
  Schematic of the products generated by HincII or HincII‐SapI digest of pICL^pt.
• F, G
  pICL^pt was replicated in mock‐ or SCAI‐depleted extracts and samples were digested with HincII (F) or HincII and SapI (G) and analyzed by agarose gel electrophoresis. DSB repair by HR restores the SapI site (Long et al, ²⁰¹¹). pQuant is used as a recovery control (Knipscheer et al, ²⁰¹²) and the quantification of this experiment is shown in Fig 3H.
• H
  pICL^pt was replicated in mock‐ or SCAI‐N‐depleted egg extracts in the presence of [α‐³²P]dATP. Samples were digested with AflIII and analyzed on a denaturing polyacrylamide gel. Stalling points relative to the ICL site are indicated. See Fig 3I for schematic of intermediates and extension products generated by AflIII digest of pICL^pt.
• I
  Samples from (H) were digested with PsiI and XhoI and analyzed on a denaturing polyacrylamide gel. Stalling points relative to the ICL site are indicated. See Fig 3I for schematic of intermediates and extension products generated by PsiI and XhoI double digest of pICL^pt.