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. 2020 Oct 30;6(44):eabb3154. doi: 10.1126/sciadv.abb3154

Fig. 2. Targeting CDK4/6 decreased the methylation of histones mediated by SMYD2 in RCTE cells and SMYD2 also regulated the expression of CDK4 and CDK6.

Fig. 2

(A to D) Western blot analysis indicated that knockdown of CDK4 and CDK6 with siRNAs (A) or knockdown of SMYD2 with siRNA (B) as well as inhibition of CDK4/6 with Abe (C) or inhibition of SMYD2 with AZ505 (D) decreased the mono-, di-, and trimethylation of histone H3 at lysine 4 (H3K4) and lysine 36 (H3K36) in RCTE cells. The quantification and statistical analysis (n = 3) were shown in the graph (bottom). *P < 0.01 as compared to each control. (E and F) Knockdown (E) or inhibition (F) of SMYD2 decreased the mRNA and protein levels of CDK4 and CDK6 in RCTE cells, examined with qRT-PCR and Western blotting. *P < 0.01 as compared to each control (n = 3). ns, not significant. (G) SMYD2 bound to the promoter of CDK4 and CDK6. ChIP-qPCR analysis was performed with an SMYD2 antibody, or normal rabbit IgG in RCTE cells. (H) ChIP assay was performed with mono-, di-, and trimethylated H3K4 and H3K36 antibodies and normal rabbit IgG in RCTE cells.