Journal: The Journal of Biological Chemistry

Article Title: O-GlcNAcylation of the tumor suppressor LATS1 drives mitotic progression via PLK1

doi: 10.1016/j.jbc.2025.110990

Figure Lengend Snippet: O-GlcNAcylation of LATS1 regulates phosphorylation of MYPT1 and PLK1. A , 293T cells were co-transfected with LATS1-WT or LATS1-4A plasmids together with HA-MYPT1, followed by treatment with nocodazole (noc). Cell lysates were subject to immunoprecipitation using an anti-HA antibody, followed by immunoblotting with the indicated antibodies. B , quantification of ( A ). ∗∗∗∗ indicates a statistically significant difference by one-way ANOVA (n = 3; ∗∗∗∗, p < 0.0001). C , 293T cells transfected with LATS1-WT or LATS1-4A were lysed and analyzed by immunoblotting the antibodies indicated. D , Quantification of ( C ). ∗∗∗∗ indicates a statistically significant difference by one-way ANOVA (n = 3; ∗∗∗∗, p < 0.0001). E , histones were extracted from 293T cells expressing LATS1-WT or LATS1-4A and immunoblotted with anti–H3-pS10 antibody. F , quantification of H3-pS10 levels in ( E ). ∗∗∗ indicates a statistically significant difference by one-way ANOVA (n = 3; ∗∗∗, p < 0.001).

Article Snippet: The following antibodies were used: anti-Myc (PTM Bio, # PTM-5390), anti-RL2 (Abcam, #ab2739), anti-GST (Gene Script, # A00865), anti-HA (DIA-An, # 3063), anti-OGT (Abcam, # ab96718), anti-LATS1 (Cell Signaling Technology, # 3477S), anti-PLK1 (Santa Cruz Biotechnology, # SC-17783), anti-H3-pS10 (Bethyl, #A301-844A), anti-H3 (Abcam, #ab4729), anti-β-actin (Sigma, Cat.# A5441), and anti-PLK1-pT210 (Abcam, Cat.# ab39068).

Techniques: Phospho-proteomics, Transfection, Immunoprecipitation, Western Blot, Expressing