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. Author manuscript; available in PMC: 2022 Aug 10.
Published in final edited form as: Dev Cell. 2021 Jul 12;56(15):2207–2222.e7. doi: 10.1016/j.devcel.2021.06.010

Figure 4. ICL-induced PIDDosome signaling does not strictly require FA pathway function.

Figure 4.

(A) Simplified diagram of the FA pathway. FACC, Fanconi core complex.

(B-C) HeLa cells transfected with indicated siRNAs were treated with or without Gö6976 and MMC (1 μM each), harvested 24hrs post-treatment and analyzed by western blot.

(D) Patient-derived GM6914 FANCA–/– fibroblasts reconstituted with empty vector or WT FANCA were treated with Gö6976 and MMC (1 μM each), harvested 24 hrs post-MMC and analyzed by western blot.

(E) HCT116 cells of indicated FANCI genotypes and stably expressing shp53 were reconstituted with empty vector (e.v) or indicated Flag-FANCI constructs and treated with or without Gö6976 (1 μM) and MMC (0.1 μM). AlamarBlue was added 72 hrs post-treatment. Data are means +/− SD of 3 independent experiments. *p < 0.05; **p < 0.01 ***p < 0.001, two-tailed Student’s t-test.

(F) Western blot from the experiment in (E) showing equal, near-physiologic expression of Flag-FANCI constructs.

(G) HeLa cells synchronized as in Table S1A were treated with or without Gö6976 and MMC (1 μM each) and harvested after 9hrs. FANCI IPs were analyzed by western blot.

(H) Schematic highlighting the effects of Chk1 (top) and Chk1i (bottom) on binding partner choice by FANCI in response to ICLs.