Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Excessive DAO inhibits myoblast migration, leading to impaired myotube fusion and muscle strength decline by reducing ECM

doi: 10.3389/fbioe.2025.1606357

Figure Lengend Snippet: DAO content in acute damaged skeletal muscle model. (A) Experimental design for glycerol-induced acute injury in 4-month-old male C57 mice. Glycerol or PBS was injected into the hind limbs. (B) Tissue appearance 3 days post-injection showed marked inflammation at glycerol injection sites. (C) HE staining of injected tissue revealed histological evidence of inflammation and tissue damage (red boxes). (D) qPCR analysis of each group (n = 3) in three times techinal repliates showed increased Aoc1 mRNA levels in glycerol-injected tissues compared to PBS-injected controls ( ***p < 0.001).

Article Snippet: Cells were treated with gradient concentrations (0, 100, 200, 500 pg/mL) of recombinant human AOC1 protein (#HY-P7497, MCE, Shanghai, China) for 24 h∼3 days.

Techniques: Injection, Staining

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Excessive DAO inhibits myoblast migration, leading to impaired myotube fusion and muscle strength decline by reducing ECM

doi: 10.3389/fbioe.2025.1606357

Figure Lengend Snippet: AOC1’s impact on myoblast migration and fusion via the Fbln1/FAK pathway. (A) Functional enrichment analysis of proteins binding to AOC1, highlighting enrichment in cytoskeletal and ECM-related proteins. (B) Morphological changes in gradient concentration effects of AOC1 on C2C12 cell fusion, showing normal fusion at 0 pg/mL, partial fusion at 100 pg/mL, and disrupted polarity at 200 pg/mL and above. (C) Immigration of cell from the upper chamber to down part was showed after AOC1 treatment, visualized by Crystal purple staining. (D) Western blot analysis confirming increased AOC1 expression and decreased Fbln1 and FAK phosphorylation (P-FAK -Y576/Y577) at higher AOC1 addtion. (E) Co-immunoprecipitation (IP) confirming AOC1’s interaction with Fbln1.

Article Snippet: Cells were treated with gradient concentrations (0, 100, 200, 500 pg/mL) of recombinant human AOC1 protein (#HY-P7497, MCE, Shanghai, China) for 24 h∼3 days.

Techniques: Migration, Functional Assay, Binding Assay, Concentration Assay, Staining, Western Blot, Expressing, Phospho-proteomics, Immunoprecipitation

Journal: Cancer Management and Research

Article Title: AOC1 Contributes to Tumor Progression by Promoting the AKT and EMT Pathways in Gastric Cancer

doi: 10.2147/CMAR.S225229

Figure Lengend Snippet: AOC1 is highly expressed in gastric cancer tissues compared with normal tissues. ( A ) The red and gray boxes indicate gastric cancer and normal tissues, respectively. Data were obtained from the GEPIA website, including 408 gastric cancer samples and 211 controls. The y-axis indicated the log2-transformed gene expression levels. qPCR ( B ) and western blot ( C ) were performed to detect AOC1 expression in the tumor and paracancerous tissues of 30 patients with gastric cancerthe. *P<0.05; **P<0.01; ***P<0.001.

Article Snippet: Primary antibodies against AOC1 (Cat#16338-1-AP, 1:1000), GAPDH (Cat# 60004-1-Ig, 1:10,000), Bax (Cat# 50599-2-Ig, 1:6000), Bcl2 (Cat# 12789-1-AP, 1:1000), Caspase-9 (Cat# 10380-1-AP, 1:300), Caspase-3 (Cat# 19677-1-AP, 1:10,000), AKT (Cat# 60203-2-Ig, 1:5000), p-AKT (Cat# 66444-1-Ig, 1:2000), Cyclin D1 (Cat# 60186-1-Ig, 1:10,000), p70S6K (Cat# 66638-1-Ig, 1:3000), E-cadherin (Cat# 60335-1-Ig, 1:5000), N-cadherin (Cat# 66219-1-Ig, 1:5000) and SNAIL (Cat# 26183-1-AP, 1:1000) were purchased from ProteinTech (Rosemont, IL, USA).

Techniques: Transformation Assay, Gene Expression, Western Blot, Expressing

Journal: Cancer Management and Research

Article Title: AOC1 Contributes to Tumor Progression by Promoting the AKT and EMT Pathways in Gastric Cancer

doi: 10.2147/CMAR.S225229

Figure Lengend Snippet: AOC1 expression is effectively knocked down by siRNA transfection in human gastric cancer cells. qRT-PCR assay was used to detect the interference efficiencies of siRNAs targeting AOC1 on mRNA levels in human ( A ) AGS and ( B ) MKN45 cells. ( C ) Western blot validated the effectiveness of AOC1 knockdown on protein levels. A scrambled siRNA was used as the negative control (NC). All experiments were performed in triplicate, independently. *P<0.05.

Article Snippet: Primary antibodies against AOC1 (Cat#16338-1-AP, 1:1000), GAPDH (Cat# 60004-1-Ig, 1:10,000), Bax (Cat# 50599-2-Ig, 1:6000), Bcl2 (Cat# 12789-1-AP, 1:1000), Caspase-9 (Cat# 10380-1-AP, 1:300), Caspase-3 (Cat# 19677-1-AP, 1:10,000), AKT (Cat# 60203-2-Ig, 1:5000), p-AKT (Cat# 66444-1-Ig, 1:2000), Cyclin D1 (Cat# 60186-1-Ig, 1:10,000), p70S6K (Cat# 66638-1-Ig, 1:3000), E-cadherin (Cat# 60335-1-Ig, 1:5000), N-cadherin (Cat# 66219-1-Ig, 1:5000) and SNAIL (Cat# 26183-1-AP, 1:1000) were purchased from ProteinTech (Rosemont, IL, USA).

Techniques: Expressing, Transfection, Quantitative RT-PCR, Western Blot, Knockdown, Negative Control

Journal: Cancer Management and Research

Article Title: AOC1 Contributes to Tumor Progression by Promoting the AKT and EMT Pathways in Gastric Cancer

doi: 10.2147/CMAR.S225229

Figure Lengend Snippet: AOC1 knockdown induces growth inhibition in human gastric cancer cells. ( A ) and ( B ) After transfection with siNC or si-AOC1, the viability of AGS and MKN45 cells was detected by using CCK-8 assay. ( C ) and ( D ) Clone formation ability of AOC1 silenced gastric cancer cells was detected. All experiments were performed in triplicate. *P<0.05.

Article Snippet: Primary antibodies against AOC1 (Cat#16338-1-AP, 1:1000), GAPDH (Cat# 60004-1-Ig, 1:10,000), Bax (Cat# 50599-2-Ig, 1:6000), Bcl2 (Cat# 12789-1-AP, 1:1000), Caspase-9 (Cat# 10380-1-AP, 1:300), Caspase-3 (Cat# 19677-1-AP, 1:10,000), AKT (Cat# 60203-2-Ig, 1:5000), p-AKT (Cat# 66444-1-Ig, 1:2000), Cyclin D1 (Cat# 60186-1-Ig, 1:10,000), p70S6K (Cat# 66638-1-Ig, 1:3000), E-cadherin (Cat# 60335-1-Ig, 1:5000), N-cadherin (Cat# 66219-1-Ig, 1:5000) and SNAIL (Cat# 26183-1-AP, 1:1000) were purchased from ProteinTech (Rosemont, IL, USA).

Techniques: Knockdown, Inhibition, Transfection, CCK-8 Assay

Journal: Cancer Management and Research

Article Title: AOC1 Contributes to Tumor Progression by Promoting the AKT and EMT Pathways in Gastric Cancer

doi: 10.2147/CMAR.S225229

Figure Lengend Snippet: Knockdown of AOC1 inhibits cell invasion and migration in human gastric cancer cells. ( A ) and ( C ) Transwell assays detecting the invasion and migration of human AGS and MKN45 cells. ( B ) and ( D ) Invaded and migrated cells in si-AOC1 group or NC group were counted. All experiments were performed for three repeated times. *P<0.05.

Article Snippet: Primary antibodies against AOC1 (Cat#16338-1-AP, 1:1000), GAPDH (Cat# 60004-1-Ig, 1:10,000), Bax (Cat# 50599-2-Ig, 1:6000), Bcl2 (Cat# 12789-1-AP, 1:1000), Caspase-9 (Cat# 10380-1-AP, 1:300), Caspase-3 (Cat# 19677-1-AP, 1:10,000), AKT (Cat# 60203-2-Ig, 1:5000), p-AKT (Cat# 66444-1-Ig, 1:2000), Cyclin D1 (Cat# 60186-1-Ig, 1:10,000), p70S6K (Cat# 66638-1-Ig, 1:3000), E-cadherin (Cat# 60335-1-Ig, 1:5000), N-cadherin (Cat# 66219-1-Ig, 1:5000) and SNAIL (Cat# 26183-1-AP, 1:1000) were purchased from ProteinTech (Rosemont, IL, USA).

Techniques: Knockdown, Migration

Journal: Cancer Management and Research

Article Title: AOC1 Contributes to Tumor Progression by Promoting the AKT and EMT Pathways in Gastric Cancer

doi: 10.2147/CMAR.S225229

Figure Lengend Snippet: Knockdown of AOC1 induces apoptosis in human gastric cancer cells. ( A ) and ( B ) The effect of AOC1 knockdown on the apoptosis of AGS cells was detected using flow cytometry. ( C ) and ( D ) The effect of AOC1 knockdown on the apoptosis of MKN45 cells was detected using flow cytometry. All experiments were performed in triplicate. *P<0.05.

Article Snippet: Primary antibodies against AOC1 (Cat#16338-1-AP, 1:1000), GAPDH (Cat# 60004-1-Ig, 1:10,000), Bax (Cat# 50599-2-Ig, 1:6000), Bcl2 (Cat# 12789-1-AP, 1:1000), Caspase-9 (Cat# 10380-1-AP, 1:300), Caspase-3 (Cat# 19677-1-AP, 1:10,000), AKT (Cat# 60203-2-Ig, 1:5000), p-AKT (Cat# 66444-1-Ig, 1:2000), Cyclin D1 (Cat# 60186-1-Ig, 1:10,000), p70S6K (Cat# 66638-1-Ig, 1:3000), E-cadherin (Cat# 60335-1-Ig, 1:5000), N-cadherin (Cat# 66219-1-Ig, 1:5000) and SNAIL (Cat# 26183-1-AP, 1:1000) were purchased from ProteinTech (Rosemont, IL, USA).

Techniques: Knockdown, Flow Cytometry

Journal: Cancer Management and Research

Article Title: AOC1 Contributes to Tumor Progression by Promoting the AKT and EMT Pathways in Gastric Cancer

doi: 10.2147/CMAR.S225229

Figure Lengend Snippet: Knockdown of AOC1 induces activation of the mitochondrial apoptosis pathway, and also inhibits the AKT signaling pathway and EMT process. ( A and B ) The key members of the mitochondrial apoptosis pathway, including Bax, Bcl2, Caspase-9, and Caspase-3, were detected by Western blot. ( C and D ) AKT signaling pathway members, including AKT, p-AKT, Cyclin D1, and p70S6K, were detected by Western blot. ( E and F ) EMT-related proteins, including E-cadherin, N-cadherin, SNAIL and Slug, were detected by Western blot. All experiments were performed in triplicate. *P<0.05.

Article Snippet: Primary antibodies against AOC1 (Cat#16338-1-AP, 1:1000), GAPDH (Cat# 60004-1-Ig, 1:10,000), Bax (Cat# 50599-2-Ig, 1:6000), Bcl2 (Cat# 12789-1-AP, 1:1000), Caspase-9 (Cat# 10380-1-AP, 1:300), Caspase-3 (Cat# 19677-1-AP, 1:10,000), AKT (Cat# 60203-2-Ig, 1:5000), p-AKT (Cat# 66444-1-Ig, 1:2000), Cyclin D1 (Cat# 60186-1-Ig, 1:10,000), p70S6K (Cat# 66638-1-Ig, 1:3000), E-cadherin (Cat# 60335-1-Ig, 1:5000), N-cadherin (Cat# 66219-1-Ig, 1:5000) and SNAIL (Cat# 26183-1-AP, 1:1000) were purchased from ProteinTech (Rosemont, IL, USA).

Techniques: Knockdown, Activation Assay, Western Blot

Journal: Cancer Management and Research

Article Title: AOC1 Contributes to Tumor Progression by Promoting the AKT and EMT Pathways in Gastric Cancer

doi: 10.2147/CMAR.S225229

Figure Lengend Snippet: IGF-1 blocked the inhibition of cell proliferation, migration and invasion caused by AOC1 knockdown. AOC1 low expressing cells were treated with IGF-1, an agonist of the AKT pathway. ( A ) CCK8 was performed to detected the proliferation of each group cells. ( B ) Transwell was performed to detected the migration and invasion of each group cells. *P<0.05 vs. NC group; #P<0.05 vs. si-AOC1 group.

Article Snippet: Primary antibodies against AOC1 (Cat#16338-1-AP, 1:1000), GAPDH (Cat# 60004-1-Ig, 1:10,000), Bax (Cat# 50599-2-Ig, 1:6000), Bcl2 (Cat# 12789-1-AP, 1:1000), Caspase-9 (Cat# 10380-1-AP, 1:300), Caspase-3 (Cat# 19677-1-AP, 1:10,000), AKT (Cat# 60203-2-Ig, 1:5000), p-AKT (Cat# 66444-1-Ig, 1:2000), Cyclin D1 (Cat# 60186-1-Ig, 1:10,000), p70S6K (Cat# 66638-1-Ig, 1:3000), E-cadherin (Cat# 60335-1-Ig, 1:5000), N-cadherin (Cat# 66219-1-Ig, 1:5000) and SNAIL (Cat# 26183-1-AP, 1:1000) were purchased from ProteinTech (Rosemont, IL, USA).

Techniques: Inhibition, Migration, Knockdown, Expressing