IndraLab

"Antibodies that recognized these phosphorylation residues were produced to study USP24 phosphorylation."

"Phosphorylation of USP24 increases its degradation during tumorigenesis and cell cycle progression."

"Phosphorylation of USP24 at these three residues was increased during mitosis (Figures 7E and F). Moreover, cdc20 could also interact with USP24 when these residues were phosphorylated, implying that the decrease in USP24 during mitosis might be important for cell cycle progression and cancer cell proliferation (Figure 7G)."

"In addition, we found that USP24 phosphorylation at Ser2604 was declined by U0126, which inhibited the Erk1/2 activities in A549 cells (Figure 7J); however, USP24 phosphorylation was not decreased by LY294002, which inhibited the PI3K activities (Supplementary Figure 5F)."

"Our previous liquid chromatography–tandem mass spectrometry (LC/MS/MS) studies probed several phosphorylation residues within USP24 (Supplementary Table 1). The phosphorylation residues were mutated individually; then, we investigated the expression of USP24-wt and its mutants (Figures 7B and C; Supplementary Figure 5B)."

"Mutations at these residues increased the USP24 half-life during mitosis, suggesting that USP24 phosphorylation during mitosis decreased USP24 expression ( xref )."

"Antibodies that recognized these phosphorylation residues were produced to study USP24 phosphorylation."

"Phosphorylation of USP24 at these three residues was increased during mitosis ( xref )."

"Three mutated residues and triple mutant S3A increased the USP24 level, suggesting that USP24 phosphorylation might be the reason for the decreased USP24 expression during mitosis and tumorigenesis."

"Our previous liquid chromatography–tandem mass spectrometry (LC/MS/MS) studies probed several phosphorylation residues within USP24 (Supplementary Table 1). The phosphorylation residues were mutated individually; then, we investigated the expression of USP24-wt and its mutants (Figures 7B and C; Supplementary Figure 5B). Three mutated residues and triple mutant S3A increased the USP24 level, suggesting that USP24 phosphorylation might be the reason for the decreased USP24 expression during mitosis and tumorigenesis. Mutations at these residues increased the USP24 half-life during mitosis, suggesting that USP24 phosphorylation during mitosis decreased USP24 expression (Figure 7D). Antibodies that recognized these phosphorylation residues were produced to study USP24 phosphorylation."

"Our previous liquid chromatography–tandem mass spectrometry (LC/MS/MS) studies probed several phosphorylation residues within USP24 (Supplementary Table 1)."

"Mutations at these residues increased the USP24 half-life during mitosis, suggesting that USP24 phosphorylation during mitosis decreased USP24 expression (Figure 7D)."

"We found that USP24 interacted with CDK1/cyclin B during the mitotic stage, and that the USP24 phosphorylation signal was higher in mitosis than in interphase ( xref ; xref )."

"Three mutated residues and triple mutant S3A increased the USP24 level, suggesting that USP24 phosphorylation might be the reason for the decreased USP24 expression during mitosis and tumorigenesis."

"Phosphorylation of USP24 at these three residues was increased during mitosis (Figures 7E and F)."

"USP24 phosphorylation increased the recruitment of the E3 ligase cdc20, which is the subunit of the APC/C complex that provides the E3 ligase activity."

"Here we found that the phosphorylation of several USP24 residues was regulated by EGF treatment and mitotic CDK1 to enhance USP24 degradation in a polyubiquitination-dependent manner."

"In addition, we found that USP24 phosphorylation at Ser2604 was declined by U0126, which inhibited the Erk1/2 activities in A549 cells ( xref ); however, USP24 phosphorylation was not decreased by LY294002, which inhibited the PI3K activities ( xref )."

"In this study, we found that the modification of USP24 by phosphorylation decreased its protein stability in KrasG12D- and EDFRL858R-induced lung cancer mouse models, lung cancer cell lines and mitotic cells."

"Here we found that the phosphorylation of several USP24 residues was regulated by EGF treatment and mitotic CDK1 to enhance USP24 degradation in a polyubiquitination-dependent manner."

"USP24 phosphorylation increased the recruitment of the E3 ligase cdc20, which is the subunit of the APC/C complex that provides the E3 ligase activity."

"Phosphorylation of USP24 increases its degradation during tumorigenesis and cell cycle progression."

"In addition to the phosphorylation of USP24 in mitosis, USP24 is also phosphorylated by EGF-activated Erk1/2 kinase activity to turnover USP24."

"Several phosphorylation residues in USP24 are involved in the control of protein stability by increasing the interaction between USP24 and the APC/C complex, suggesting that APC/C might be important for the downregulation of USP24."

"In addition to the phosphorylation of USP24 in mitosis, USP24 is also phosphorylated by EGF-activated Erk1/2 kinase activity to turnover USP24."

"Regulation of USP24 by phosphorylation increases its degradation."

"(B) Various mutations of USP24 phosphorylation residues, including a triple mutant (S3A), were constructed and expressed in cells. Samples were analyzed by western blotting with an anti-GFP antibody (a). The GFP-USP24 level was quantified after three independent experiments (b). (C) The proteins stabilities of USP24-wt (WT) and its mutants (S1616A, S2047A, S2604A and S3A) were investigated after cycloheximide (CHX) treatment. Samples were analyzed by western blotting with an anti-GFP antibody (a). Proteins levels were quantified after three independent experiments (b). (D) Normal or mitotic A549 cells were collected for an immunoblotting assay with antibodies against USP24, phospho-USP24 (S1616), phospho-USP24 (S2047), phospho-USP24 (S2604), cyclin B1 and actin. (E, F) Samples were collected under nocodazole treatment (NZ) (E) or nocodazole release (NZ-R) (F). Samples were analyzed with antibodies against USP24 phosphorylation at indicated residues, cyclin B1 and actin. (G) A549 cells treated with nocodazole were collected for an immunoprecipitation assay with anti-cdc20. The immunoprecipitated samples were used for western blotting with antibodies against the indicated proteins. (H) Samples collected from EGF-treated mice lung primary cells were analyzed by western blotting with antibodies against indicated proteins. (I) Samples were collected from A549 cells treated with FTI-276, and then analyzed by western blotting with antibodies against USP24 phosphorylation at the indicated residues (a). The relative levels were quantified after three independent experiments (b). (J) Samples were collected from A549 cells treated with Erk inhibitor, and then analyzed by western blotting with antibodies against USP24 phosphorylation at the indicated residues (a)."

"(B) Various mutations of USP24 phosphorylation residues, including a triple mutant (S3A), were constructed and expressed in cells. Samples were analyzed by western blotting with an anti-GFP antibody (a). The GFP-USP24 level was quantified after three independent experiments (b). (C) The proteins stabilities of USP24-wt (WT) and its mutants (S1616A, S2047A, S2604A and S3A) were investigated after cycloheximide (CHX) treatment. Samples were analyzed by western blotting with an anti-GFP antibody (a). Proteins levels were quantified after three independent experiments (b). (D) Normal or mitotic A549 cells were collected for an immunoblotting assay with antibodies against USP24, phospho-USP24 (S1616), phospho-USP24 (S2047), phospho-USP24 (S2604), cyclin B1 and actin. (E, F) Samples were collected under nocodazole treatment (NZ) (E) or nocodazole release (NZ-R) (F). Samples were analyzed with antibodies against USP24 phosphorylation at indicated residues, cyclin B1 and actin. (G) A549 cells treated with nocodazole were collected for an immunoprecipitation assay with anti-cdc20. The immunoprecipitated samples were used for western blotting with antibodies against the indicated proteins. (H) Samples collected from EGF-treated mice lung primary cells were analyzed by western blotting with antibodies against indicated proteins. (I) Samples were collected from A549 cells treated with FTI-276, and then analyzed by western blotting with antibodies against USP24 phosphorylation at the indicated residues (a)."