We are sorry, but NCBI web applications do not support your browser and may not function properly. More information

Results: 3

1.
FIGURE 2.

FIGURE 2. From: HDAC6 Is Required for Epidermal Growth Factor-induced β-Catenin Nuclear Localization.

HDAC6 deacetylates β-catenin at Lys-49 and inhibits phosphorylation of β-catenin at Ser-45. a, immunoblot analysis of HCT116 cells. Cells were serum-starved overnight and treated with EGF for the indicated times, and β-catenin was immunoprecipitated (IP) followed by Western blot with antibodies to β-catenin and HDAC6. b, immunoblot analysis of HCT116 cells transfected with HDAC6 siRNA. 48 h after transfection, lysates were immunoprecipitated with β-catenin antibody and immunoblotted with acetylated lysine antibody (AcK). Con, control. c, alignment of N-terminal amino acid sequences of β-catenin. d, immunoblot analysis of Myc immunoprecipitates from 293T cells expressing Myc-β-catenin or Myc-β-catenin (K49R), using antibodies to Ac-β-cat (Lys-49) and p-β-cat (Ser-45). e, immunoblot analysis of HCT116 cells expressing HA-CBP, wild-type (wt) FLAG-HDAC6, or catalytically dead (mut) FLAG-HDAC6. f, immunoblot analysis of HCT116 and SW480 cells transfected with HDAC6 siRNA or treated with TSA. Cells were serum-starved overnight and treated with EGF for 4 h. g, immunoblot analysis of 293T cells expressing Myc-β-catenin, together with FLAG-HDAC6. h, immunoblot analysis of SW480 cells expressing FLAG-HDAC6. 16 h after transfection, cells were serum-starved overnight before cell lysates were collected.

Yu Li, et al. J Biol Chem. 2008 May 9;283(19):12686-12690.
2.
FIGURE 3.

FIGURE 3. From: HDAC6 Is Required for Epidermal Growth Factor-induced β-Catenin Nuclear Localization.

HDAC6 inactivation blocks EGF-induced β-catenin nuclear localization and results in growth inhibition. a, immunoblot analysis of HCT116 cells. Cells were serum-starved overnight and treated with EGF for the indicated times followed by immunoprecipitation (IP) with antibody to caveolin-1. b, immunoblot analysis of HCT 116 cells. Cells were serum-starved overnight and treated with cyclodextrin (CD, 5 mm for 2 h) before EGF treatment (30 min) followed by immunoprecipitation with antibody to caveolin-1. c, immunoblotting of nuclear fraction for β-catenin in HCT116 cells transfected with HDAC6 and β-catenin siRNAs. 36 h after siRNA transfection, cells were serum-starved overnight and treated with EGF for 3 h. The intensity of signals of Ac-β-cat (Lys-49) was quantified, normalized to total β-catenin, and displayed in graphical format. All results are representative of three independent experiments. con, control. d, viable cell number after siRNA transfection. Cells were collected at 48 h after transfection, stained with trypan blue, and counted. Results are representative of three independent experiments.

Yu Li, et al. J Biol Chem. 2008 May 9;283(19):12686-12690.
3.
FIGURE 1.

FIGURE 1. From: HDAC6 Is Required for Epidermal Growth Factor-induced β-Catenin Nuclear Localization.

EGF-induced β-catenin nuclear localization is inhibited by TSA. a, immunoblot analysis of HCT116 cells transfected with β-catenin siRNA. Cells were cultured in Dulbecco’s modified Eagle’s medium with 10% fetal bovine serum, and cell lysates were made 48 h after transfection. con, control. b, immunoblot analysis of HCT116 and SW480 cells treated with TSA. Cells were serum-starved overnight and treated with EGF for 4 h or treated with TSA for 8 h before EGF treatment. Cell lysates were immunoprecipitated (IP) with β-catenin and immunoblotted with acetylated lysine antibody (AcK). Ac-tubulin (Ac-tub) was used as a control for TSA treatment. c, immunofluorescence staining for β-catenin in HCT116 cells. Cells were serum-starved overnight and treated with EGF for 3 h or treated with TSA for 8 h before EGF treatment. All nuclei were stained with 4,6-diamidino-2-phenylindole (DAPI). d, immunoblotting of nuclear and cytosolic fractions for β-catenin in HCT116 described in panel c. Lamin A was used as a nuclear marker.

Yu Li, et al. J Biol Chem. 2008 May 9;283(19):12686-12690.

Supplemental Content

Recent activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...
Write to the Help Desk