Fig. 4: Lactylation of NUDT21 facilitates NUDT21–CPSF6 complex formation.
From: NUDT21 lactylation reprograms alternative polyadenylation to promote cuproptosis resistance
a, b Immunoblot analysis of the interaction between NUDT21 and CPSF6 in KYSE30 cells treated with 20 mM l-lactate (a) or 20 mM oxamate (b). c Immunoblot analysis of the interaction between NUDT21 and CPSF6 in KYSE30 cells transfected with AARS1-HA, with or without β-alanine (100 mM) stimulation for 24 h. d Immunoblot analysis of the interaction between NUDT21 and CPSF6 in KYSE30 cells transfected with siRNA specific for HDAC2. e Immunoblot analysis of the interaction between NUDT21 and CPSF6 in KYSE30 cells treated with acetate (10 mM) or l-lactate (20 mM). Cells were pre-treated with oxamate. f In vitro pull-down assays detecting the interaction between CPSF6 and NUDT21. Left: schematic of purification. Cells were pre-treated with oxamate before l-lactate or acetate treatment. g Immunoblot analysis of the interaction between NUDT21 and its binding partners in KYSE30 cells transfected with the indicated plasmids. h Experimental schematic of the PLA assay. i, j Representative images of PLA foci showing the interaction between NUDT21 and CPSF6 in KYSE30 cells treated with saline, l-lactate (20 mM), or oxamate (20 mM). Scale bars, 5 μm. The right panel is quantification, n = 50 cells. k–m RIP assays assessing the association between NUDT21 and FDX1 RNA in KYSE30 cells under the indicated conditions. Data are presented as mean ± SD; P value was calculated by Student’s t-test (i–m).
