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. Author manuscript; available in PMC: 2017 Aug 1.
Published in final edited form as: Cell Rep. 2017 Jun 20;19(12):2432–2440. doi: 10.1016/j.celrep.2017.05.081

Figure 3. Ubiquitination-Independent and Ubiquitination-Linked FANCI Phosphorylation.

Figure 3

(A) Immunoblot of FANCA-deficient (GM6914), FANCF-deficient (TOV21G) and FANCG-deficient (326SV) cells, and their complemented counterparts, that were either untreated (Utx), or treated with 60 ng/mL MMC for 24 hours.

(B) Immunoblot of FANCD2-deficient (PD20F) cells, complemented with either wild-type FANCD2 (FANCD2wt) or a monoubiquitination-deficient (K561R) mutant, that were either untreated, or treated with 60 ng/mL MMC for 24 hours.

(C) Immunoblot of pellet (chromatin containing) fractions prepared from FANCI-deficient (F010191) cells complemented with either wild-type FANCI (FANCIwt) or a monoubiquitination-deficient (K523R) mutant, that were either untreated, or treated with 60 ng/mL MMC for 24 hours. Irrelevant lanes have been removed from these blots.

(D) Immunoblot of FANCI-deficient (HCT116 FANCI−/−) cells complemented with an empty vector, wild-type FANCI, or the indicated mutants. Cells were then either untreated, or treated with 60 ng/mL MMC for 24 hours. Ratios of monoubiquitinated to non-ubiquitinated FANCD2 (D2 Ub:non-Ub) are indicated beneath the corresponding lanes.

Ub refers to the monoubiquitinated forms of FANCI and FANCD2, and non-Ub refers to the non-ubiquitinated forms. The asterisk (*) indicates a non-specific band.

See also Figures S3 and S4.