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. Author manuscript; available in PMC: 2018 Feb 16.
Published in final edited form as: Mol Cell. 2017 Feb 2;65(4):699–714.e6. doi: 10.1016/j.molcel.2017.01.008

Figure 7. hUBE2T-deficient RA2627 cells expressing hUBE2T S5D phosphomimetic are defective for FANCD2 and FANCI monoubiquitination.

Figure 7

(A) Anti-HA western blot of hUBE2T deficient transformed and immortalized RA2627 E6/E7/hTERT fibroblasts expressing the indicated protein. These cells were used in panels B-D.

(B) Western blot analysis of FANCD2 and FANCI monoubiquitination in hUBE2T complemented cells following 24hr treatment with 1µM MMC.

(C) FANCD2 foci formation in hUBE2T complemented cells following 24hr treatment with 1µM MMC.

(D) Quantification of FANCD2 foci formation visualized in C.

(E) MMC cell survival assay of hUBE2T complemented cells, see Figure S6C for expression levels.

(F) Western blot of HA-FLAG-tagged hUBE2T expression in immortalized RA2627 cells used in G-H.

(G) Quantification of FANCD2 foci formation of hUBE2T S5A mutant compared to EV and WT expressing RA2627 fibroblasts, see Figure S6E for visualization.

(H) MMC cell survival assay of hUBE2T complemented cells.

See also Figure S6.