Journal: International Journal of Molecular Sciences

Article Title: Dual Oxidase 2 (DUOX2) as a Proteomic Biomarker for Predicting Treatment Response to Chemoradiation Therapy for Locally Advanced Rectal Cancer: Using High-Throughput Proteomic Analysis and Machine Learning Algorithm

doi: 10.3390/ijms232112923

Figure Lengend Snippet: ( A ) Machine learning approaches to measure the predictive power of proteins for CCRT response. ( B ) Immunohistochemical (IHC) validation of dual oxidase 2 (DUOX2) as a predictive biomarker for CCRT response (statistical significance, * p -value < 0.05).

Article Snippet: The monoclonal mouse anti-DUOX2 antibody (NB110-61576, Novus Biologicals, Centennial, CO, USA) was diluted to 1:100.

Techniques: Immunohistochemical staining, Biomarker Discovery

Journal: PLoS ONE

Article Title: CRISPR-Cas9 Mediated NOX4 Knockout Inhibits Cell Proliferation and Invasion in HeLa Cells

doi: 10.1371/journal.pone.0170327

Figure Lengend Snippet: ( A ) T7 endonuclease 1(T7E1) assay. HeLa cells were transiently transfected with either pX459/NOX4 gRNA #1 or pX459/NOX4 gRNA #2, and genomic PCR products were analyzed by T7E1 assay (left panel). HeLa cells transfected with pX459/NOX4 gRNA’s were selected with puromycin and single colonies were isolated. The genomic PCR product of each clone was analyzed by T7E1 assay (right panel). Experiments were repeated three times with similar observations, and representative images are shown. ( B ) DNA sequence analysis showed the presence of the NOX4 mutation in clones #1–2, #1–5 and #1–6. The black arrows indicate the heterogeneous genomic DNA sequences in each cell lines. ( C ) CRISPR-Cas9 introduced the insertion and deletion (indel) mutation in the target sites. ( D ) Western blot of NOX4 knockout cell lines. Equal amount of HeLa control and NOX4 knockout cell lysates were probed with anti-NOX4 antibody. Experiments were repeated three times with similar observations, and representative data is shown. ( E ) NOX4 knockout did not influence NOX1, NOX2, NOX3, NOX5, DUOX1 and DUOX2 levels. Protein levels of NOX1, NOX2, NOX3, NOX5, DUOX1 and DUOX2 in parental HeLa cells and three clones of NOX4 knockout cells. ( F ) NOX4 knockout showed lower H 2 O 2 production. The levels of H 2 O 2 were measured with the Amplex Red assay in triplicate. The graph shows the average and the standard deviation (SD). Control vs knockout cells. *: P <0.005, **: P <0.0001.

Article Snippet: The antibody for NOX5 from Boster Biological Technology (Pleasanton, CA), the antibody for DUOX1 from GeneTex (Irvine, CA), the antibody for DUOX2 from Novus Biologicals (Littleton, CO), and the antibody for p22phox from Santa Cruz Biotechnology (San Diego, CA).

Techniques: Transfection, Isolation, Sequencing, Mutagenesis, Clone Assay, CRISPR, Western Blot, Knock-Out, Control, Amplex Red Assay, Standard Deviation