Extended Data Fig. 3: HMCES suppresses ATR-dependent CHK1 phosphorylation during NEIL3-dependent ICL repair.
From: The HMCES DNA-protein cross-link functions as an intermediate in DNA interstrand cross-link repair
a, b, and d, HMCES immunodepletions. The extracts used in Fig. 2d (a), Extended Data Fig. 3c (b), and Extended Data Fig. 3e (d) were blotted for HMCES. Asterisks, non-specific bands. c, pICLAP was replicated in mock- or HMCES-depleted extracts supplemented with ATR inhibitor AZD6738 (ATRi), as indicated. Replication reactions were separated by SDS-PAGE and blotted for phospho-CHK1 and MCM6 (loading control). Accumulation of phosphorylated CHK1 was blocked by AZD6738 treatment, indicating that CHK1 phosphorylation is dependent on ATR. e, pCtrl, pICLPt, or pICLAP was replicated in mock- or HMCES-depleted extracts, as indicated. Replication reactions were analyzed as in c. HMCES depletion increased the accumulation of phosphorylated CHK1 only during replication of pICLAP, indicating a specific role for HMCES in NEIL3-dependent ICL repair.
