![Fig. 5. Overexpression of a dominant-negative Akt (Akt[KM]) impairs the ability of IL-3 to override γ-IR–induced cell-cycle arrest. / (A) Whole cell lysates from 32D-Neo (lane 1) or 2 clones of 32D-Akt (lanes 2 and 3) or 32D-Akt(KM) (lanes 3 and 4) cells were Western blotted with anti-HA antibodies (αHA) or anti-Akt/PKB PH domain antiserum (αAkt). (B) 32D-Neo, 32D-Akt, and 32D-Akt(KM) cells were cultured in medium that contained 70 pg/mL IL-3. Subsequently, cultures were exposed to 4 Gy γ-IR (+γ) or were left untreated (−γ). Twenty-four hours after irradiation, cells were stained with PI and analyzed by flow cytometry. Before PI staining, cell viability was determined by trypan blue exclusion and was 94% or greater in all experiments. Percentage of S-phase cells is indicated for each culture.](https://ash.silverchair-cdn.com/ash/content_public/journal/blood/98/3/10.1182_blood.v98.3.834/5/h81511366005.jpeg?Expires=1722627660&Signature=F9tOu7bk9bQbz5NHVybkK9msx11ZH5Nw0fg3wGNG889ffBk~ctSWW1MDpGZw-2FBh6uWgVCkO4D35m0Y8W6OaUT9uw~77GUNwSwAzJbvNtl6YsM1pBmHcAwPdAISqCMfwfB1WAgRDAaODeEP408WDLLv3nOtfX9LBQ-PbRKv1kgMzmpD~vv~yzFLA0xXGivctLMZYc8d0~aGHIr2972x~FNyZw8epLqifIcL771QRr8gfj~xCByI-k4Pmi-1PPf1CszTu~EpTmIZUhoJaeLpIGN9P3R6YVU2hGejcuuzP4faFAr-vhKCouPcLyccxGNDPNdWfHAlp525~wtMO3isuQ__&Key-Pair-Id=APKAIE5G5CRDK6RD3PGA)
Overexpression of a dominant-negative Akt (Akt[KM]) impairs the ability of IL-3 to override γ-IR–induced cell-cycle arrest.
(A) Whole cell lysates from 32D-Neo (lane 1) or 2 clones of 32D-Akt (lanes 2 and 3) or 32D-Akt(KM) (lanes 3 and 4) cells were Western blotted with anti-HA antibodies (αHA) or anti-Akt/PKB PH domain antiserum (αAkt). (B) 32D-Neo, 32D-Akt, and 32D-Akt(KM) cells were cultured in medium that contained 70 pg/mL IL-3. Subsequently, cultures were exposed to 4 Gy γ-IR (+γ) or were left untreated (−γ). Twenty-four hours after irradiation, cells were stained with PI and analyzed by flow cytometry. Before PI staining, cell viability was determined by trypan blue exclusion and was 94% or greater in all experiments. Percentage of S-phase cells is indicated for each culture.
