Acetylation of FAM134B K160 is responsive to ER stress. (A) The reliability of FAM134B K160 acetylation antibody was validated by performing a dot blot assay using synthesized acetyl-peptide (C)WEVINSK(Ac)PDER-NH2 (amino acids 154–164, NP_001030023.1) and the control unacetylated peptide (C)WEVINSKPDER-NH2. (B) K160 acetylation of FAM134B in FAM134B-Flag stable cell line. IP was performed with an antibody to acetylated lysine. The control was the Protein A/G agarose resin without the antibody that can recognize acetylated FAM134B. K160 acetylation and Flag were detected by Western blot (WB). (C and D) ER stress markers including ATF6, p-IRE1, and p-PERK were response to EBSS or Tg treatment. HEK293T cells were treated with EBSS or Tg (1 μM) to trigger ER stress for 0, 0.5, 1 h. The indicated proteins were detected by Western blot. (E) K160 acetylation of FAM134B in FAM134B-Flag stable cell line increased by treated with deacetylase inhibitors. Cells were treated with deacetylase inhibitors TSA (1 mM) and NAM (5 mM) for 12 h. The control was treated with DMSO. IP was performed with an antibody to acetylated lysine. K160 acetylation and Flag were detected by Western blot. Molecular weight measurements are in kD. Source data are available for this figure: SourceData FS1.