amigo2 antibody  (ZenBio)

Journal: NPJ Precision Oncology

Article Title: Metabolomic and transcriptomic profiling of HNSCC identifies AMIGO2 as a therapeutic target modulating tumor microenvironment

doi: 10.1038/s41698-025-01132-z

Figure Lengend Snippet: a Expression of AMIGO2, STC2, INHBA, DKK1, VEGFC, SPOCK1, MT1E, and FOXD1 in scRNA-seq UMAP plots. b Bar plot showing cell type proportions under LRS subtypes from scRNA-seq data. c AMIGO2, ranked highly for prognostic importance, was selected for further analysis. d AMIGO2 expression is upregulated in most tumor tissues across various cancers in TCGA pan-cancer dataset. e Evaluation of AMIGO2’s prognostic value (OS) in TCGA pan-cancer dataset. f Correlation analysis between AMIGO2 expression and representative signatures in TCGA pan-cancer dataset. Ns: no significant difference, * P < 0.05, ** P ≤ 0.01, *** P ≤ 0.001.

Article Snippet: The following primary antibodies were used for Western blotting: AMIGO2 (1:500, HUAAN, catalog No: ER1903-66), GAPDH (1:1000, CST, catalog No: 5174S, RRID: AB_10622025), E-cadherin (1:1000, CST, catalog No: 3195S, RRID: AB_2291471), N-cadherin (1:1000, CST, catalog No: 13116S, RRID: AB_2687616), HPRT1 (1:1000, Proteintech Cat# 15059-1-AP, RRID: AB_10638622), PAICS (1:1000, Proteintech Cat# 12967-1-AP, RRID: AB_10638449), Vimentin (1:1000, CST, catalog No: 5741S, RRID: AB_10695459), MMP9 (1:1000, CST, catalog No: 13667S, RRID: AB_2798289), FAP (Thermo Fisher Scientific Cat# PA5-99313, RRID: AB_2818246), α-SMA (Cell Signaling Technology Cat# 19245, RRID: AB_2734735), PI3K (Cell Signaling Technology Cat# 4292, RRID:AB_329869), Phospho-PI3K (Cell Signaling Technology Cat# 17366, RRID:AB_2895293), AKT (Cell Signaling Technology Cat# 9272, RRID: AB_329827), Phospho-Akt (Thr308) (Cell Signaling Technology Cat# 9275, RRID: AB_329828), Phospho-Akt (Ser473) (Cell Signaling Technology Cat# 9271, RRID: AB_329825), PDK1 (Thermo Fisher Scientific Cat# A302-130A, RRID:AB_1720395), Phospho-PDK1 (Ser241) (Cell Signaling Technology Cat# 3061, RRID: AB_2161919), mTOR (Abcam Cat# ab134903, RRID: AB_2800465) and mTOR (phospho S2481) (Abcam Cat# ab137133, RRID:AB_2800466).

Techniques: Expressing

Journal: NPJ Precision Oncology

Article Title: Metabolomic and transcriptomic profiling of HNSCC identifies AMIGO2 as a therapeutic target modulating tumor microenvironment

doi: 10.1038/s41698-025-01132-z

Figure Lengend Snippet: a IHC images and quantitative statistics of AMIGO2 expression in adjacent normal tissue, precancerous lesions, and tumor tissues. b Representative images and quantification of CD8, CD56, FAP, and PanCK staining in tumor tissues from high-AMIGO2 and low-AMIGO2 patient groups. c Graphical abstract for this study. Ns: no significant difference. * P < 0.05, ** P ≤ 0.01, *** P ≤ 0.001.

Article Snippet: The following primary antibodies were used for Western blotting: AMIGO2 (1:500, HUAAN, catalog No: ER1903-66), GAPDH (1:1000, CST, catalog No: 5174S, RRID: AB_10622025), E-cadherin (1:1000, CST, catalog No: 3195S, RRID: AB_2291471), N-cadherin (1:1000, CST, catalog No: 13116S, RRID: AB_2687616), HPRT1 (1:1000, Proteintech Cat# 15059-1-AP, RRID: AB_10638622), PAICS (1:1000, Proteintech Cat# 12967-1-AP, RRID: AB_10638449), Vimentin (1:1000, CST, catalog No: 5741S, RRID: AB_10695459), MMP9 (1:1000, CST, catalog No: 13667S, RRID: AB_2798289), FAP (Thermo Fisher Scientific Cat# PA5-99313, RRID: AB_2818246), α-SMA (Cell Signaling Technology Cat# 19245, RRID: AB_2734735), PI3K (Cell Signaling Technology Cat# 4292, RRID:AB_329869), Phospho-PI3K (Cell Signaling Technology Cat# 17366, RRID:AB_2895293), AKT (Cell Signaling Technology Cat# 9272, RRID: AB_329827), Phospho-Akt (Thr308) (Cell Signaling Technology Cat# 9275, RRID: AB_329828), Phospho-Akt (Ser473) (Cell Signaling Technology Cat# 9271, RRID: AB_329825), PDK1 (Thermo Fisher Scientific Cat# A302-130A, RRID:AB_1720395), Phospho-PDK1 (Ser241) (Cell Signaling Technology Cat# 3061, RRID: AB_2161919), mTOR (Abcam Cat# ab134903, RRID: AB_2800465) and mTOR (phospho S2481) (Abcam Cat# ab137133, RRID:AB_2800466).

Techniques: Expressing, Staining

Journal: Scientific reports

Article Title: Prevention of liver metastasis via the pharmacological suppression of AMIGO2 expression in tumor cells.

doi: 10.1038/s41598-024-71827-z

Figure Lengend Snippet: Fig. 3. Suppression of AMIGO2 mRNA and protein expression in LV12 cells treated with trametinib, ruxolitinib, and SP600125 and their reduced adhesion to mouse liver sinusoidal endothelial (HSE) cells. (a) LV12 cells were treated with trametinib, ruxolitinib, SP600125, or DMSO for 24 h. The relative AMIGO2 mRNA levels in each drug treatment group, compared with that in DMSO-treated LV12 cells, was determined using qRT-PCR. The bar graphs show mean ± SD from three to five independent experiments (n = 9–15, Student’s t-test). (b) Immunoreactive AMIGO2 expression was quantified as a ratio to β-actin expression using densitometry. Values represent mean ± SD (n = 7, Student’s t-test) from at least six independent experiments with similar results. (c) Typical immunoblotting performed using anti-AMIGO2 is shown in the upper panel, and the lower panel presents immunoblotting using anti-β-actin antibodies. Full-length blots were presented in Supplementary Fig. S5. (d) LV12 cells treated as described in “a” were labeled with PKH67 fluorescent dye and placed on HSE cells. The percentage of adherent cells was determined by measuring the fluorescence intensity. The graph shows mean ± SD (n = 4 in each group, Student’s t-test) from four independent experiments with similar results.

Article Snippet: For LV12 cells, the membranes were incubated with mouse monoclonal anti-AMIGO2 antibody (G-7; sc-373699, Santa Cruz Biotechnology, Santa Cruz, CA, USA) at a 1:50 dilution or rabbit polyclonal anti-AMIGO2 antibody (LS-C401236; LS Bio, Shirley, MA, USA) at a 1:1,000 dilution, and then with peroxidase-conjugated goat polyclonal anti-mouse IgG antibody at a 1:2,000 dilution (PM009-7; Medical & Biological Laboratories, Nagoya, Japan) or peroxidase-conjugated goat polyclonal anti-rabbit IgG antibody at a 1:2,000 dilution (ab97080; Abcam, Cambridge, UK), respectively.

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Labeling, Fluorescence

Journal: Scientific reports

Article Title: Prevention of liver metastasis via the pharmacological suppression of AMIGO2 expression in tumor cells.

doi: 10.1038/s41598-024-71827-z

Figure Lengend Snippet: Fig. 3. Suppression of AMIGO2 mRNA and protein expression in LV12 cells treated with trametinib, ruxolitinib, and SP600125 and their reduced adhesion to mouse liver sinusoidal endothelial (HSE) cells. (a) LV12 cells were treated with trametinib, ruxolitinib, SP600125, or DMSO for 24 h. The relative AMIGO2 mRNA levels in each drug treatment group, compared with that in DMSO-treated LV12 cells, was determined using qRT-PCR. The bar graphs show mean ± SD from three to five independent experiments (n = 9–15, Student’s t-test). (b) Immunoreactive AMIGO2 expression was quantified as a ratio to β-actin expression using densitometry. Values represent mean ± SD (n = 7, Student’s t-test) from at least six independent experiments with similar results. (c) Typical immunoblotting performed using anti-AMIGO2 is shown in the upper panel, and the lower panel presents immunoblotting using anti-β-actin antibodies. Full-length blots were presented in Supplementary Fig. S5. (d) LV12 cells treated as described in “a” were labeled with PKH67 fluorescent dye and placed on HSE cells. The percentage of adherent cells was determined by measuring the fluorescence intensity. The graph shows mean ± SD (n = 4 in each group, Student’s t-test) from four independent experiments with similar results.

Article Snippet: For LV12 cells, the membranes were incubated with mouse monoclonal anti-AMIGO2 antibody (G-7; sc-373699, Santa Cruz Biotechnology, Santa Cruz, CA, USA) at a 1:50 dilution or rabbit polyclonal anti-AMIGO2 antibody (LS-C401236; LS Bio, Shirley, MA, USA) at a 1:1,000 dilution, and then with peroxidase-conjugated goat polyclonal anti-mouse IgG antibody at a 1:2,000 dilution (PM009-7; Medical & Biological Laboratories, Nagoya, Japan) or peroxidase-conjugated goat polyclonal anti-rabbit IgG antibody at a 1:2,000 dilution (ab97080; Abcam, Cambridge, UK), respectively.

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Labeling, Fluorescence