Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

Article Title: Knockdown of Dual Oxidase 1 (DUOX1) Promotes Wound Healing by Regulating Reactive Oxygen Species (ROS) by Activation of Nuclear Factor kappa B (NF-κB) Signaling

doi: 10.12659/MSM.926492

Figure Lengend Snippet: The sequences of DUOX1 siRNA (siDUOX1).

Article Snippet: The membrane was then incubated with primary antibodies against DUOX1 (1: 1000, orb589688; Biorbyt), collagen I (1: 1000, Ab138492; Abcam), collagen III (1: 1000, Ab184993; Abcam), p21 (1: 2000, Ab109520; Abcam), p16 (1: 5000, Ab51243; Abcam), NF-κB (1: 2000, Ab16502; Abcam), H3 (1: 1000, Ab1791; Abcam), and GAPDH (1: 2000, #5174; Cell Signaling Technology [CST]) overnight at 4°C with gentle shaking.

Techniques:

Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

Article Title: Knockdown of Dual Oxidase 1 (DUOX1) Promotes Wound Healing by Regulating Reactive Oxygen Species (ROS) by Activation of Nuclear Factor kappa B (NF-κB) Signaling

doi: 10.12659/MSM.926492

Figure Lengend Snippet: DUOX1 was highly expressed in slow-healing granulation tissue of burn wounds ( A ) Clinical burned wounds with fast healing (n=12) and slow healing (n=12) granulation tissue and normal tissue (n=6) were collected, and Q-PCR was used to detect the expression of DUOX1. Primary fibroblasts were isolated from fast-healing, slow-healing, and normal tissues. ( B ) Collagen I and III protein levels were detected by Western blotting. ( C ) Cell proliferation was detected by CCK8 at 0, 24, 48, and 72 h. * P <0.05, ** P <0.01, *** P <0.001 vs normal; # P <0.05, ## P <0.01 vs slow-healing. Normal tissues: skin tissue taken from the surface of traumatic injury, fast-healing granulation tissue: better recovery after 14 days’ treatment, slow-healing (n=12) granulation tissue: poor recovery after 14 days’ treatment.

Article Snippet: The membrane was then incubated with primary antibodies against DUOX1 (1: 1000, orb589688; Biorbyt), collagen I (1: 1000, Ab138492; Abcam), collagen III (1: 1000, Ab184993; Abcam), p21 (1: 2000, Ab109520; Abcam), p16 (1: 5000, Ab51243; Abcam), NF-κB (1: 2000, Ab16502; Abcam), H3 (1: 1000, Ab1791; Abcam), and GAPDH (1: 2000, #5174; Cell Signaling Technology [CST]) overnight at 4°C with gentle shaking.

Techniques: Expressing, Isolation, Western Blot

Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

Article Title: Knockdown of Dual Oxidase 1 (DUOX1) Promotes Wound Healing by Regulating Reactive Oxygen Species (ROS) by Activation of Nuclear Factor kappa B (NF-κB) Signaling

doi: 10.12659/MSM.926492

Figure Lengend Snippet: Knockdown of DUOX1 significantly increased cell proliferation and inhibited the production of reactive oxygen species (ROS). ( A ) Following primary isolation from wound granulation tissue, fibroblasts (Vimentin-positive) were identified by immunohistochemical staining. ( B, C ) Primary fibroblasts were transfected with human DUOX1 interferon, and the interference efficiency of DUOX1 was detected by Q-PCR ( B ) and Western blot ( C ). Following DUOX1 gene interference in primary fibroblasts, ( D ) a CCK-8 kit was used to detect cell proliferation; ( E, F ) biochemical detection of cell supernatant MDA ( E ) and SOD ( F ) expression; ( G, H ) flow detection of ROS and ( G ) cell apoptosis ( H ); ( I ) Western blotting of NF-κB (nuclear and plasma), collagen, collagen III, P21, and P16. ** P <0.01, *** P <0.001 vs siNC. siNC – negative control of DUOX1 interference.

Article Snippet: The membrane was then incubated with primary antibodies against DUOX1 (1: 1000, orb589688; Biorbyt), collagen I (1: 1000, Ab138492; Abcam), collagen III (1: 1000, Ab184993; Abcam), p21 (1: 2000, Ab109520; Abcam), p16 (1: 5000, Ab51243; Abcam), NF-κB (1: 2000, Ab16502; Abcam), H3 (1: 1000, Ab1791; Abcam), and GAPDH (1: 2000, #5174; Cell Signaling Technology [CST]) overnight at 4°C with gentle shaking.

Techniques: Knockdown, Isolation, Immunohistochemical staining, Staining, Transfection, Western Blot, CCK-8 Assay, Expressing, Clinical Proteomics, Negative Control

Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

Article Title: Knockdown of Dual Oxidase 1 (DUOX1) Promotes Wound Healing by Regulating Reactive Oxygen Species (ROS) by Activation of Nuclear Factor kappa B (NF-κB) Signaling

doi: 10.12659/MSM.926492

Figure Lengend Snippet: DUOX1-regulated wound healing, likely by regulation of reactive oxygen species (ROS). ( A, B ) Transfected primary fibroblasts with overexpression of human DUOX1 were constructed, and Q-PCR ( A ) and Western blot ( B ) were used to detect the efficiency of DUOX1 overexpression. Following pre-transfection of DUOX1 overexpression in primary fibroblasts for 24 h, cells were treated with NAC and ROS inhibitor (10 μM). ( C, D ) Expression of MDA ( C ) and SOD ( D ) were detected in cell supernatants; ( E, F ) flow cytometry was used to detect ROS and ( E ) cell apoptosis ( F ); ( G ) cell proliferation activity was detected using a CCK-8 kit; ( H ) expression of NF-κB (nuclear and plasma), collagen I, collagen III, P21, and P16 were detected by Western blotting. * P <0.05, ** P <0.01, *** P <0.001 vs vector or vehicle+vector; ## P <0.01, ### P <0.001 vs vehicle+oeDUOX1.

Article Snippet: The membrane was then incubated with primary antibodies against DUOX1 (1: 1000, orb589688; Biorbyt), collagen I (1: 1000, Ab138492; Abcam), collagen III (1: 1000, Ab184993; Abcam), p21 (1: 2000, Ab109520; Abcam), p16 (1: 5000, Ab51243; Abcam), NF-κB (1: 2000, Ab16502; Abcam), H3 (1: 1000, Ab1791; Abcam), and GAPDH (1: 2000, #5174; Cell Signaling Technology [CST]) overnight at 4°C with gentle shaking.

Techniques: Transfection, Over Expression, Construct, Western Blot, Expressing, Flow Cytometry, Activity Assay, CCK-8 Assay, Clinical Proteomics, Plasmid Preparation

Journal: bioRxiv

Article Title: Pharmacological CDK4/6 inhibition unravels a p53-induced secretory phenotype in senescent cells

doi: 10.1101/2020.06.05.135715

Figure Lengend Snippet: p16-3MR mice were treated with vehicle (PBS, 7 consecutive days), doxorubicin (5 mg/kg, 3 consecutive days) or abemaciclib (50 mg/kg, 7 consecutive days). N=6 mice/group. 14 dpt, bioluminescence was visualized and quantified by the IVIS spectrum in vivo imaging system, as shown by representative bioluminescence images ( A ) and quantification ( B ). ( C ) RNA isolated from treated kidneys and mRNA encoding p16 quantified by qRT-PCR. Representative images ( D ) to visualize SA-β-gal activities in vehicle-, doxorubicin-or abemaciclib-treated mouse kidney sections at 15 dpt (arrows indicated positive area; scale bar, 1 mm; N=3) and quantified ( E ). ( F ) 15 dpt, plasma was collected and expression levels of CXCL1 measured by ELISA. ( G ) Protein lysate obtained from drug treated kidneys to quantify IL6 by ELISA. ( H and I ) RNA isolated from kidneys and mRNA encoding indicated genes quantified by qRT-PCR. ( J ) Relative weight changes were calculated at 7 dpt and 14 dpt. Red blood cells ( K ) and white blood cells ( L ) were counted at 15 dpt. Percentage of T cells, B cells, granulocytes and macrophages were determined by flow cytometry analysis ( M ). Physical performance was measured by rotarods assay at 15 dpt ( N ), grip strength meter at 7 dpt and 14 dpt ( O ), and hanging tests were performed at 7 dpt and 14 dpt and normalized to weights ( P ). One-way ANOVA, data are means ±SD ( B, C, E, F, G, K, L and N ). Two-way ANOVA, data are means ±SD ( H, I, J, M, O and P ). *p<0.05, **p<0.01, ***p<0.001, N.S.=not significant. dpt, days post treatment.

Article Snippet: 20 μg total protein was collected from each treated kidney and IL6 protein level was determined by the mouse IL6 duo-set (R&D Systems) ELISA.

Techniques: In Vivo Imaging, Isolation, Quantitative RT-PCR, Clinical Proteomics, Expressing, Enzyme-linked Immunosorbent Assay, Flow Cytometry