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. Author manuscript; available in PMC: 2021 Apr 30.
Published in final edited form as: Cell. 2020 Apr 20;181(3):702–715.e20. doi: 10.1016/j.cell.2020.03.051

Figure 5. Phosphorylation of Ser241 Is Necessary and Sufficient for Functional Activity of MYBL2.

Figure 5.

(A) Cell cycle effects of MYBL2 shRNA knockdown with simultaneous overexpression of WT MYBL2, mutant MYBL2 S241A (nonphosphorylatable alanine mutant), S241D (a phosphomimetic mutation), or TAD-deleted MYBL2 (TAD_del) were examined in KOPT-K1 cells.

(B) Mean percentages of cells ± SD are shown in each phase of the cell cycle and sub-G1 (apoptotic) cells for the DNA histograms in (A). *p < 0.05 and **p < 0.01 by Student’s t test for 3 biological replicates versus controls.

(C) Sensitivity to iHAP1 in MYBL2 knockdown KOPT-K1 cells conditionally expressing the phosphomimetic S241D mutant or WT MYBL2 as mean ± SD of three biological replicates. Control shRNA targets luciferase (sh_Luc). *p < 0.05, **p < 0.01, and ***p < 0.001 by Student’s t test.

(D) The MYBL2 shRNA rescue experiment included simultaneous overexpression in KOPT-K1 cells of WT MYBL2 or S241A, S241D, or TAD_del mutant MYBL2 constructs. Acetocarmine, Alexa Fluor 647 (red) anti-a tubulin antibody, and DAPI were used to stain chromatin, microtubules, and DNA, respectively.