Fig. 4.

Interaction of ESCO2 with hnRNPA1 acetylates hnRNPA1 at K277. a ESCO2-FLAG vector was transfected into HEK293T cells, the ESCO2-FLAG complexes underwent Co-IP, and then ESCO2-binding proteins were identified by combined silver staining and MS. b Enrichment plot showing enrichment of spliceosome-related genes in the ESCO2 high-expression group in TCGA LUAD cohort. c Heatmap showing the relative expression values for 126 spliceosome-related genes in TCGA LUAD cohort. d The ESCO2-FLAG vector was transfected into NCI-H1975 cells, the ESCO2-FLAG complexes underwent Co-IP, and then hnRNPA1 in the complexes was detected. e hnRNPA1-HAplasmids were transfected into NCI-H1975 cells, the hnRNPA1-HA complexes underwent Co-IP, and then ESCO2 in the complexes was detected. f NCI-H1975 cells were simultaneously transfected with hnRNPA1-HAand ESCO2-FLAGvectors, immunoprecipitated with anti-HA antibody, and then detection was performed using anti–ac-K antibody. g NCI-H1975 cells with ESCO2 stable silencing were transfected with hnRNPA1-HA vector, immunoprecipitated by anti-HA antibody, and then detection was performed using anti–ac-K antibody. h The K277 acetylation site was identified by MS. i hnRNPA1 WT or K277R mutant plasmids were co-transfected with ESCO2-FLAG into NCI-H1975 cells, and acetylation levels were detected using anti–ac-K antibody. j and k hnRNPA1 WT or K277R mutant plasmids were co-transfected with ESCO2-FLAG into NCI-H1975 cells, anti-HA antibody was used for Co-IP, and ESCO2-FLAG was detected using anti-FLAG antibody (j); anti-FLAG antibody was used for Co-IP, and hnRNPA1 WT and K277R mutant constructs were detected using anti-HA antibody (k). Recombinant WT hnRNPA1 and its mutant K277R were incubated with recombinant ESCO2, and acetylation levels were detected using anti–ac-K antibody