![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=1735594276&Signature=W7se0dRRAqq38LaR0gTros6270R2Cq29UUvfDWLXI7pvhr0iej9E08XwEmnc9-s6-KxaQno2aoIusc6MuTGfTrmGA~HK-WmVHSr~t0rc93rn~od4FsYo2~35S0UIocqJkRoYSm-1z8gaZxZi-K9BUZHOpnPMal9a~J9YXHEniUHwT1-OIiZnC87o84EeYzlKB58POj4KQj-5hL9RJxOE5VK4prsrcpHl2Oa1J4~W7UWrUji7s7F3RfEOubnnEBxi0tlJWP9LIArwwm-Xg9fFRVGXNj0guSNAIdJKguN0MV1OpAlK7DOFRzQklmVDMeNrau-a6LQr-z9VerDcfbW4hg__&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.
