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. Author manuscript; available in PMC: 2020 Mar 18.
Published in final edited form as: Cancer Cell. 2019 Feb 28;35(3):504–518.e7. doi: 10.1016/j.ccell.2019.01.020

Figure 4. Regulation of DNA damage response by phosphorylation of PTEN tyrosine 240 is independent of its lipid phosphatase activity.

Figure 4.

(A) U87-WT/YF/GR-PTEN cells were examined for pY240 by western blot. (B) Cells from (A) were treated with 10 Gy IR and analyzed for pH2AX at indicated times. (C and D) Cells from (A) were treated with 10 Gy IR and analyzed by immunofluorescence at the indicated times for the presence DSBs. Quantification of γH2AX (C) and RAD51 (D) foci positive cells is shown. (E) Cells treated as (C) were analyzed for colony formation. (F) U87-WT/YF/GR-PTEN cells were examined for AKT phosphorylation at Ser473 by western blot. (G) Proposed therapeutic strategies for GBM patients according to PTEN genetic status and phosphorylation of Y240. (H) HK281 cells reconstituted with WT-, YF-, or GR/YF-PTEN were treated with or without 3 Gy IR and analyzed for survival by limiting dilution assay. (I) U87-WT/YF/GR- or G129R/Y240F (GR/YF)-PTEN cells along with FGFR2 were treated with 10 Gy IR and chromatin-bound protein isolated at indicated times post-IR was immunoblotted for PTEN and RAD51. Error bars in this figure are SD of three independent experiments. *p<0.05. See also Figure S3.