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. Author manuscript; available in PMC: 2021 Nov 5.
Published in final edited form as: Mol Cell. 2020 Oct 26;80(3):410–422.e6. doi: 10.1016/j.molcel.2020.10.008

Figure 3. Depletion of FAM122A promotes WEE1 stability in A549 lung cancer cells.

Figure 3.

(A) Western blots of the lysates from control and FAM122A-KO A549 cells after treatment with prexasertib (100nM) for 24hrs.

(B) Western blots of the lysates from control and FAM122A-KO A549 cells treated with/without prexasertib (100nM) and MG132 for 16 hours. The relative levels of phosphorylated WEE1 compare to WEE1 were quantified as indicated.

(C) (Upper panels) Western blots of the lysates from control and FAM122A-KO A549 cells treated with DMSO or prexasertib (100nM) for 24 hrs, followed by treatment with cycloheximide (CHX) for the indicated duration. (Bottom panel) Quantification of WEE1 protein levels. Error bars indicate standard errors (n=3).

(D) Western blots of the lysates from control and FAM122A-KO A549 cells treated with prexasertib (100nM) or AZD1775 (50nM) for 24 hrs.

(E) Survival plots of control and FAM122A-KO A549 cells treated with graded concentrations of prexasertib along with 50nM WEE1i (AZD1775). Note that WEE1i sensitizes resistant FAM122A-KO A549 cells to prexasertib. Data are shown as mean ± SD from three independent experiments. sgFAM122A versus sgFAM122A+AZD1775, ****P<0.0001, statistical analysis was performed using two-way ANOVA.