Fig. S1

Autophagy recruitment at stalled replication forks promotes TOP1cc degradation, related to Figure 1 and Table S1 (A) Co-immunoprecipitation of endogenous TEX264 in WT cells or in the context of ATG7 depletion (n = 2). Unspecific immunoglobulin G (IgG) was used instead of TEX264 antibody as a control. (B) Overlap of the protein set identified by immunoprecipitation of replication forks interactome by MS64,65,66 and known autophagy-related proteins from the human UniProt-Swissprot database, the Gene Ontology (GO) database, and the Kyoto Encyclopedia of Genes and Genomes (KEGG) database. Replisomes were identified by iPOND, or Bio-ID of PCNA, in different cell lines after treatment with different replication stress inducers. HU, hydroxyurea; UV, ultraviolet. (C) LysoIP was performed in HeLa WT or cells modified for LysoIP experiments by expression of the resident lysosomal tagged protein TMEM192-HA (n = 3). The LysoIP cell line allows the specific pull-down of lysosome vesicles without contamination from other cellular organelles. (D) Immunofluorescence of HeLa WT cells and TMEM192-HA cells used for LysoIP, using anti-HA antibody and anti-LAMP1 marker of the late endosome and lysosome vesicles (n = 3). Scale bar, 10 μm. (E) LysoIP performed after 6 h of treatment with 50 nM CPT in S-phase-synchronized HCT116 cells (n = 3). All conditions were treated with 50 nM BAF. (F) LysoIP performed after 6 h of treatment with 50 nM CPT in S-phase-synchronized cells in hTERT RPE-1 (n = 3). All conditions were treated with 50 nM BAF. (G) LysoIP performed in HeLa released from a single thymidine block into the S-phase and treated for 3 h with 50 nM CPT, 5 μM irinotecan, or 50 nM SN38 (n = 3). All conditions are treated with 50 nM BAF.