Inhibition of S151 phosphorylation does not affect K160 acetylation. (A) Measurement of interaction between HA-CBP and FAM134B (WT)-Flag, FAM134B (S151A)-Flag, and FAM134B (S151D)-Flag. IP was performed with anti-HA beads, which was followed by Western blot (WB) for FAM134B-Flag. (B) Measurement of interaction between Myc-SIRT7 and FAM134B (WT)-Flag, FAM134B (S151A)-Flag, and FAM134B (S151D)-Flag. IP was performed with anti-Myc magnetic beads, which was followed by Western blot for FAM134B-Flag. (C and D) K160 acetylation was not affected by expression of S151D mimicking permanent phosphorylation. HEK293T cells transfected with FAM134B (WT)-Flag, FAM134B (S151A)-Flag, and FAM134B (S151D)-Flag were treated with Tg (1 μM) for 0, 0.5, 1 h. IP was performed with anti-Flag beads, which was followed by Western blot for K160 acetylation. Quantification of K160 acetylation was shown in D (n = 3 experimental replicates). Data are shown as means ± SEM and analyzed with one-way ANOVA. **P < 0.01, ***P < 0.001, ns means no significance. (E) K160 acetylation was not affected by CAMK2B KD. CAMK2B WT or KD cells were treated with Tg (1 μM) for 1 h or DMSO. Cells were collected and analyzed for proteins as indicated by Western blot. Molecular weight measurements are in kD. Source data are available for this figure: SourceData FS4.