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. 2022 Jul 7;16(15):2771–2787. doi: 10.1002/1878-0261.13264

Fig. 5.

Fig. 5

p27 is the key SKP2 substrate that defines CHK1i sensitivity. (A) Baf3 SKP2 KO clones were transduced with two independent sg‐p27/dsRed or sg‐Cd8a/dsRed constructs, respectively. Three days post transduction the bulks were treated with PF‐477736 [1 μm] or DMSO for 48 h. The percentage of dsRed+ cells was assessed using flow cytometry, whereby treated cells were normalized to DMSO controls. Bars indicate the mean percentage (±SEM) of two independent clones analyzed in three independent experiments. (B) Western blot showing p27 levels in Baf3 p27 KO clones, Skp2 KO, and parental Baf3 cells. (C) Baf3 p27‐KO and sg‐Cd8a control clones were transduced with two different sg‐Skp2/dsRed constructs. Three days post transduction the cells were treated with CHK1 inhibitor PF‐477736 [1 μm] for 48 h. The percentage of dsRed+ cells was assessed using flow cytometry, whereby treated cells were normalized to DMSO controls. Bars indicate the mean percentage (±SEM) of two independent clones per genotype in three independent experiments.