Journal: Oncology Reports

Article Title: Knockdown of HIP1 expression promotes ligand-induced endocytosis of EGFR in HeLa cells

doi: 10.3892/or.2017.6025

Figure Lengend Snippet: EGFR expression in seventeen cell lines measured by western blotting. The cell lines were 7402, 7703, 7721, Hep3B, HepG2, H460, SPCA1, SKOV-3, HeLa, MCF-7, glioma cells U87, U251, C33a, PC-3, NCI-H1299, NCI-H446, and K562 cells. Hep3B, HeLa, PC-3, NCI-H1299, and NCI-H466 cells exhibited higher expression levels of EGFR than those of other cell lines. Combined with the results presented in , HeLa cells had high expression of both HIP1 and EGFR.

Article Snippet: After SDS-PAGE, the protein was transferred to a nitrocellulose membrane using a semidry transfer apparatus (Bio-Rad, Hercules, CA, USA), blocked with 3% bovine serum albumin in TBST [10 mmol/l Tris (pH 8.0), 150 mmol/l NaCl, 0.1% Tween-20], incubated at room temperature for 1 h with rabbit anti-human HIP1, EGFR, and GAPDH antibodies, rotated for 1 h at room temperature of 25°C, and washed with TBST three times.

Techniques: Expressing, Western Blot

Journal: Oncology Reports

Article Title: Knockdown of HIP1 expression promotes ligand-induced endocytosis of EGFR in HeLa cells

doi: 10.3892/or.2017.6025

Figure Lengend Snippet: Expression levels of EGFR in HeLa cells with HIP1 Stealth RNAi. Neither siRNA1 nor siRNA2 had an effect on the expression levels of EGFR and GAPDH in HeLa cells.

Article Snippet: After SDS-PAGE, the protein was transferred to a nitrocellulose membrane using a semidry transfer apparatus (Bio-Rad, Hercules, CA, USA), blocked with 3% bovine serum albumin in TBST [10 mmol/l Tris (pH 8.0), 150 mmol/l NaCl, 0.1% Tween-20], incubated at room temperature for 1 h with rabbit anti-human HIP1, EGFR, and GAPDH antibodies, rotated for 1 h at room temperature of 25°C, and washed with TBST three times.

Techniques: Expressing

Journal: Oncology Reports

Article Title: Knockdown of HIP1 expression promotes ligand-induced endocytosis of EGFR in HeLa cells

doi: 10.3892/or.2017.6025

Figure Lengend Snippet: Expression levels of EGFR in NCI-H1299 cells after HIP1 Stealth RNAi. siRNAs had no effect on the expression of EGFR or GAPDH in NCI-H1299 cells.

Article Snippet: After SDS-PAGE, the protein was transferred to a nitrocellulose membrane using a semidry transfer apparatus (Bio-Rad, Hercules, CA, USA), blocked with 3% bovine serum albumin in TBST [10 mmol/l Tris (pH 8.0), 150 mmol/l NaCl, 0.1% Tween-20], incubated at room temperature for 1 h with rabbit anti-human HIP1, EGFR, and GAPDH antibodies, rotated for 1 h at room temperature of 25°C, and washed with TBST three times.

Techniques: Expressing

Journal: Oncology Reports

Article Title: Knockdown of HIP1 expression promotes ligand-induced endocytosis of EGFR in HeLa cells

doi: 10.3892/or.2017.6025

Figure Lengend Snippet: Endocytosis of EGFR and localization of clathrin in HeLa and HeLa HIP1 RNAi2 cells (1.5 ng/ml EGF stimulation). After 1.5 ng/ml EGF stimulation, endocytosis of EGF-bound EGFR and clathrin were accelerated after the expression of HIP1 was blocked. EGFR and clathrin were colocalized in the cytoplasm.

Article Snippet: After SDS-PAGE, the protein was transferred to a nitrocellulose membrane using a semidry transfer apparatus (Bio-Rad, Hercules, CA, USA), blocked with 3% bovine serum albumin in TBST [10 mmol/l Tris (pH 8.0), 150 mmol/l NaCl, 0.1% Tween-20], incubated at room temperature for 1 h with rabbit anti-human HIP1, EGFR, and GAPDH antibodies, rotated for 1 h at room temperature of 25°C, and washed with TBST three times.

Techniques: Expressing

Journal: Oncology Reports

Article Title: Knockdown of HIP1 expression promotes ligand-induced endocytosis of EGFR in HeLa cells

doi: 10.3892/or.2017.6025

Figure Lengend Snippet: Endocytosis of EGFR and localization of clathrin in HeLa and HeLa HIP1 RNAi2 cells (100 ng/ml EGF stimulation). After 100 ng/ml EGF stimulation, endocytosis of EGF-bound EGFR and clathrin were accelerated significantly after the expression of HIP1 was blocked. EGFR and clathrin were colocalized in the cytoplasm.

Article Snippet: After SDS-PAGE, the protein was transferred to a nitrocellulose membrane using a semidry transfer apparatus (Bio-Rad, Hercules, CA, USA), blocked with 3% bovine serum albumin in TBST [10 mmol/l Tris (pH 8.0), 150 mmol/l NaCl, 0.1% Tween-20], incubated at room temperature for 1 h with rabbit anti-human HIP1, EGFR, and GAPDH antibodies, rotated for 1 h at room temperature of 25°C, and washed with TBST three times.

Techniques: Expressing