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. 2021 Aug 24;12(1):220–235. doi: 10.1158/2159-8290.CD-21-0560

Figure 3.

Figure 3. DNMT3A is degraded through the ubiquitin-proteasome system. A, Survival after the birth of mice with the indicated genotypes. No Dnmt3a−/−, Dnmt3aW293Del/−, or Dnmt3aW293Del/W293Del mice survived beyond postnatal day 24. B, Western blot of DNMT3A and GAPDH protein expression in whole bone marrow cells of mice. C, Gene expression analysis of Dnmt3a in whole bone marrow measured by quantitative PCR and normalized for Gapdh mRNA expression. D, The stability ratio of the WT and W297Del mutant over the indicated times, calculated as described in Fig. 2C. E, Alterations in DNMT3A protein stability after administration of proteasome inhibitor (MG132), a CRL inhibitor (MLN4924), an autophagy inhibitor (chloroquine), and an unfolded protein response inhibitor (IREi). F, Stability ratio of MFI of DNMT3A-GFP versus MFI of DsRed before and after treatment with the proteasome inhibitor (MG132) as measured by flow cytometry 48 hours after transfection. Data are presented as mean ± SEM. *, P < 0.05; **, P < 0.01; ***, P < 0.001 using the unpaired t test. See also Supplementary Fig. S7.

DNMT3A is degraded through the ubiquitin-proteasome system. A, Survival after the birth of mice with the indicated genotypes. No Dnmt3a−/−, Dnmt3aW293Del/−, or Dnmt3aW293Del/W293Del mice survived beyond postnatal day 24. B, Western blot of DNMT3A and GAPDH protein expression in whole bone marrow cells of mice. C, Gene expression analysis of Dnmt3a in whole bone marrow measured by quantitative PCR and normalized for Gapdh mRNA expression. D, The stability ratio of the WT and W297Del mutant over the indicated times, calculated as described in Fig. 2C. E, Alterations in DNMT3A protein stability after administration of proteasome inhibitor (MG132), a CRL inhibitor (MLN4924), an autophagy inhibitor (chloroquine), and an unfolded protein response inhibitor (IREi). F, Stability ratio of MFI of DNMT3A-GFP versus MFI of DsRed before and after treatment with the proteasome inhibitor (MG132) as measured by flow cytometry 48 hours after transfection. Data are presented as mean ± SEM. *, P < 0.05; **, P < 0.01; ***, P < 0.001 using the unpaired t test. See also Supplementary Fig. S7.