rabbit polyclonal anti hyou1  (Cell Signaling Technology Inc)

Journal: Molecules and Cells

Article Title: Expression of HYOU1 via Reciprocal Crosstalk between NSCLC Cells and HUVECs Control Cancer Progression and Chemoresistance in Tumor Spheroids

doi: 10.14348/molcells.2020.0212

Figure Lengend Snippet: (A) A gene expression heat map representing fold changes greater than 1.5 in samples from lung cancer spheroids grown in CM from cultured HUVECs, cultured NCI-H460 cells, or lung cancer cells co-cultured with HUVECs. (B and C) Categorization of biological pathways (B) and biological processes (C) identified by microarray analysis as markedly altered in lung cancer spheroids grown in CM from cultured HUVECs, cultured NCI-H460 cells, or lung cancer cells co-cultured with HUVECs. (D) Expression levels of CARS, HYOU1, MARS, and pAKT (Ser437) in lung cancer spheroids grown in CM from NCI-H460 cells cultured alone or co-cultured with HUVECs, as assessed by western blot analysis.

Article Snippet: After washing with DPBS three times, the spheroids were incubated with rabbit polyclonal anti-HYOU1 (1:100; Cell Signaling Technology, USA) in DPBS with 10% normal goat serum (Vector Laboratories, USA) for 16 h at 4°C, and then washed three times for 10 min with DPBS.

Techniques: Expressing, Cell Culture, Microarray, Western Blot

Journal: Molecules and Cells

Article Title: Expression of HYOU1 via Reciprocal Crosstalk between NSCLC Cells and HUVECs Control Cancer Progression and Chemoresistance in Tumor Spheroids

doi: 10.14348/molcells.2020.0212

Figure Lengend Snippet: (A) Bright-field images of 3D spheroids co-cultured with HUVECs and NSCLC cells (NCI-H460 or A549) and spheroids cultured with lung cancer cells alone. The images were obtained using the Operetta ® High Content Screening System with a 10× objective. (B) Expression levels of ATF6, HYOU1, IRE1, and pAKT (Ser473) in spheroids co-cultured with HUVECs and NSCLC cells (NCI-H460 or A549) and spheroids cultured with lung cancer cells alone, as assessed by western blot analysis. (C) Multilayer image showing immunofluorescence staining of HYOU1 in NSCLC (NCI-H460 or A549) spheroids co-cultured with HUVECs and Hoechst staining of both cell types. (D) Expression levels of cleaved caspase-3 and HYOU1 in lung cancer cells transfected with nonspecific siRNA (siCont) or HYOU1 siRNA (siHYOU1) and co-cultured with HUVECs with or without 10 μM or 20 μM gefitinib or cisplatin for 72 h, as assessed by western blot analysis.

Article Snippet: After washing with DPBS three times, the spheroids were incubated with rabbit polyclonal anti-HYOU1 (1:100; Cell Signaling Technology, USA) in DPBS with 10% normal goat serum (Vector Laboratories, USA) for 16 h at 4°C, and then washed three times for 10 min with DPBS.

Techniques: Cell Culture, High Content Screening, Expressing, Western Blot, Immunofluorescence, Staining, Transfection

Journal: Molecules and Cells

Article Title: Expression of HYOU1 via Reciprocal Crosstalk between NSCLC Cells and HUVECs Control Cancer Progression and Chemoresistance in Tumor Spheroids

doi: 10.14348/molcells.2020.0212

Figure Lengend Snippet: (A) Expression levels of HIF1 and HYOU1 in monolayer (2D)- or spheroid (3D)-cultured NSCLC cells (NCI-H460, A549, H1299, and PC9), as assessed by western blot analysis. (B) Clonogenic survival in NSCLC cells (NCI-H460, A549, and H1299) transfected with nonspecific siRNA (siCont) or HYOU1 siRNA (siHYOU1), as assessed by colony formation assay. (C) Immunofluorescence and bright-field images of lung cancer spheroids (NCI-H460, A549, and H1299) transfected with nonspecific siRNA (siCont) or HYOU1 siRNA (siHYOU1). The spheroids were stained with 4 μM EthD-1. The images were obtained using the Operetta ® High Content Screening System, and the intensity of EthD-1 staining in lung cancer spheroids relative to controls was analyzed using Harmony software. (D) Expression of cleaved caspase-3, cleaved PARP, HIF1, HYOU1, pp38, p53, and pErk1/2 in lung cancer spheroids (NCI-H460 and A549) transfected with nonspecific siRNA (siCont) or HYOU1 siRNA (siHYOU1). The data shown are the mean ± SD from three independent experiments; ** P < 0.01, *** P < 0.001, and **** P < 0.0001 compared to the control group.

Article Snippet: After washing with DPBS three times, the spheroids were incubated with rabbit polyclonal anti-HYOU1 (1:100; Cell Signaling Technology, USA) in DPBS with 10% normal goat serum (Vector Laboratories, USA) for 16 h at 4°C, and then washed three times for 10 min with DPBS.

Techniques: Expressing, Cell Culture, Western Blot, Transfection, Colony Assay, Immunofluorescence, Staining, High Content Screening, Software

Journal: Molecules and Cells

Article Title: Expression of HYOU1 via Reciprocal Crosstalk between NSCLC Cells and HUVECs Control Cancer Progression and Chemoresistance in Tumor Spheroids

doi: 10.14348/molcells.2020.0212

Figure Lengend Snippet: (A) CSC spheroid formation of H1299, A549, and NCI-H460 cells transfected with nonspecific siRNA (siCont) or HYOU1 siRNA (siHYOU1). (B) Expression levels of CD133 and HYOU1 in H1299 cells transfected with nonspecific siRNA (siCont) or HYOU1 siRNA (siHYOU1). (C) Bright-field and immunofluorescence images of lung cancer spheroids (NCI-H460 and A549) transfected with nonspecific siRNA (siCont) or HYOU1 siRNA (siHYOU1) and treated with 20 μM gefitinib. The spheroids were stained with 4 μM EthD-1. The images were obtained using the Operetta ® High Content Screening System, and the intensity of EthD-1 staining in lung cancer spheroids was analyzed using Harmony software. (D) Bright-field images of NSCLC cells (NCI-H460 and A549) transfected with nonspecific siRNA (siCont) or HYOU1 siRNA (siHYOU1), as assessed by migration assay. (E) Expression levels of α-SMA, collagen I, HYOU1, N-cadherin, and vimentin in in lung cancer spheroids (NCI-H460 and A549) transfected with nonspecific siRNA (siCont) or HYOU1 siRNA (siHYOU1). The data shown are the mean ± SD from three independent experiments; * P < 0.05 compared to the control group.

Article Snippet: After washing with DPBS three times, the spheroids were incubated with rabbit polyclonal anti-HYOU1 (1:100; Cell Signaling Technology, USA) in DPBS with 10% normal goat serum (Vector Laboratories, USA) for 16 h at 4°C, and then washed three times for 10 min with DPBS.

Techniques: Transfection, Expressing, Immunofluorescence, Staining, High Content Screening, Software, Migration

Journal: Molecules and Cells

Article Title: Expression of HYOU1 via Reciprocal Crosstalk between NSCLC Cells and HUVECs Control Cancer Progression and Chemoresistance in Tumor Spheroids

doi: 10.14348/molcells.2020.0212

Figure Lengend Snippet: (A) Target pathways identified by microarray analysis as markedly altered in NCI-H460 cells transfected with siHYOU1. (B) Expression levels of IFN-α, and IFN-β in NSCLC cells (NCI-H460 and A549) transfected with nonspecific siRNA (siCont) or HYOU1 siRNA (siHYOU1).

Article Snippet: After washing with DPBS three times, the spheroids were incubated with rabbit polyclonal anti-HYOU1 (1:100; Cell Signaling Technology, USA) in DPBS with 10% normal goat serum (Vector Laboratories, USA) for 16 h at 4°C, and then washed three times for 10 min with DPBS.

Techniques: Microarray, Transfection, Expressing

Journal: Molecules and Cells

Article Title: Expression of HYOU1 via Reciprocal Crosstalk between NSCLC Cells and HUVECs Control Cancer Progression and Chemoresistance in Tumor Spheroids

doi: 10.14348/molcells.2020.0212

Figure Lengend Snippet: (A) Expression levels of HYOU1 in NSCLC cells (NCI-H460 and H1299) treated with 0.1 μM or 1 μM of an mTOR inhibitor (Torin2 or WYE-125132) or a PI3K inhibitor (GDC0032 or PKI-402). (B) Expression levels of HYOU1 and mTOR in NSCLC cells (NCI-H460 and H1299) transfected with nonspecific siRNA (siCont), HYOU1 siRNA (siHYOU1), or mTOR siRNA (simTOR). The data shown are the mean ± SD from three independent experiments; * P < 0.05 compared to the control group.

Article Snippet: After washing with DPBS three times, the spheroids were incubated with rabbit polyclonal anti-HYOU1 (1:100; Cell Signaling Technology, USA) in DPBS with 10% normal goat serum (Vector Laboratories, USA) for 16 h at 4°C, and then washed three times for 10 min with DPBS.

Techniques: Expressing, Transfection