Journal: Materials today. Bio

Article Title: Precise delivery of doxorubicin and imiquimod through pH-responsive tumor microenvironment-active targeting micelles for chemo- and immunotherapy.

doi: 10.1016/j.mtbio.2022.100482

Figure Lengend Snippet: Fig. 10. Immunostaining of tumor tissues in tumor-bearing mice after treatments. (A) Immunohistochemistry images of tumor sections stained with CD3, CD8, and TNF-α antibodies. The scale bar is 100 μm. (B) Immunofluorescence images of tumor tissues after treatments for 12 days. The scale bar is 50 μm. Blue fluorescence represents the cell nucleus stained with DAPI. Green fluorescence represents the iNOS stained with the iNOS antibody conjugated with FITC.

Article Snippet: After 30 min, the tissue slice was stained with diluted iNOS antibody (Miltenyi Biotec, catalog: 130-116-357) at 4 C overnight.

Techniques: Immunostaining, Immunohistochemistry, Staining

Journal: Acta Pharmacologica Sinica

Article Title: Efficacy of subantimicrobial-dose doxycycline against nitrosative stress in chronic periodontitis

doi: 10.1038/aps.2012.129

Figure Lengend Snippet: Photomicrographs of representative iNOS-positive and 3-nitrotyrosine-positive stains in gingival tissue from patients with chronic periodontitis (immunohistochemical staining): (A) countless iNOS+ PMNs and monocytes (scale bar, 50 μm); (B) decreased iNOS+ cells in the SRP+placebo group (scale bar, 50 μm); (C) sparse iNOS+ cells in SRP+SDD group (scale bar, 50 μm); (D) countless 3NT+ PMNs and monocytes (scale bar, 100 μm); (E) decreased 3NT+ cells in the SRP+placebo group (scale bar, 50 μm); (F) sparse 3NT+ cells in the SRP+SDD group (scale bar, 50 μm). This figure is representative of 3 measurements (×400). iNOS+=induced nitric oxide synthase positive cells; 3NT+=3-nitrotyrosine positive cells; SRP=scaling and root planing; SDD=subantimicrobial dose of doxycycline.

Article Snippet: The following primary monoclonal antibodies were used: anti iNOS (R&D Systems, RD-MAB9502, MN, USA), anti 3-nitrotyrosine (3NT) (Novus Europe, KA0445, ABNOVA, Cambridge, UK).

Techniques: Immunohistochemical staining, Staining

Journal: Acta Pharmacologica Sinica

Article Title: Efficacy of subantimicrobial-dose doxycycline against nitrosative stress in chronic periodontitis

doi: 10.1038/aps.2012.129

Figure Lengend Snippet: Periodontal immunohystochemical and serum nitrosative stress parameters of the patients.

Article Snippet: The following primary monoclonal antibodies were used: anti iNOS (R&D Systems, RD-MAB9502, MN, USA), anti 3-nitrotyrosine (3NT) (Novus Europe, KA0445, ABNOVA, Cambridge, UK).

Techniques:

Journal: Cell reports

Article Title: Proinflammatory signaling in islet β cells propagates invasion of pathogenic immune cells in autoimmune diabetes

doi: 10.1016/j.celrep.2022.111011

Figure Lengend Snippet:

Article Snippet: anti-iNOS , Fluidigm , 3161011C.

Techniques: Recombinant, Plasmid Preparation, Blocking Assay, Red Blood Cell Lysis, Reverse Transcription, Cell Isolation, Staining, RNA Sequencing, Software

Journal: Oncology Letters

Article Title: Overexpression of MTH1 and OGG1 proteins in ulcerative colitis-associated carcinogenesis

doi: 10.3892/ol.2018.8812

Figure Lengend Snippet: Antibodies used in immunohistochemical stain.

Article Snippet: Finally, sections were counterstained with Mayer's hematoxylin. table ft1 table-wrap mode="anchored" t5 Table I. caption a7 Antigen Primary antibodies Dilution Source Antigen retrieval iNOS nos typeII Mouse monoclonal 1:100 BD Biosciences, Franklin Lakes, NJ Microwaved for 30 min in target retrieval solution high pH (DAKO, Glostrup, Denmark) 8-OHdG N45.1 Mouse monoclonal 1:20 Japanese aging control institute, Shizuoka, Japan Proteinase K for 7 min incubate with 4N HCl for 20 min and with 50 mM Tris base for 5 min OGG1 NB100-106 Rabbit polyclonal 1:250 Novus Biologicals, Littleton, CO, Microwaved for 30 min in target retrieval solution high pH (DAKO, Glostrup, Denmark) MTH1 D-2 Rabbit polyclonal 1:400 Abcam, Cambridge, England Microwaved for 30 min in target retrieval solution high pH (DAKO, Glostrup, Denmark) MUTYH ab13698 Rabbit-polyclonal 1:50 Abcam, Cambridge, England Microwaved for 30 min in target retrieval solution high pH (DAKO, Glostrup, Denmark) p53 PAb1801 Mouse monoclonal 1:100 Oncogene Research Products, San Diego, CA.

Techniques: Immunohistochemical staining, Staining

Journal: Nitric oxide : biology and chemistry

Article Title: NITRIC OXIDE-MEDIATED RESISTANCE TO PHOTODYNAMIC THERAPY IN A HUMAN BREAST TUMOR XENOGRAFT MODEL: IMPROVED OUTCOME WITH NOS2 INHIBITORS

doi: 10.1016/j.niox.2016.12.003

Figure Lengend Snippet: Western blot depicting iNOS upregulation in ALA/light-challenged MDA-MB-232 cells. Cells in serum-free medium were dark-incubated with 1 mM ALA for 30 min, switched to ALA-free medium, and exposed to a 1 J/cm2 fluence of broad-band visible light. Immediately after irradiation (0 h) and after increasing periods of subsequent dark incubation up to 24 h, cells were retrieved for Western analysis of iNOS and β-actin. A dark control (DC: ALA alone for 24 h) was also analyzed. Number below each iNOS band indicates band intensity relative to β-actin and normalized to DC. Total protein load per lane: 100 μg.

Article Snippet: The polyclonal antibody against human iNOS was obtained from Cayman Chemicals (Ann Arbor, MI).

Techniques: Western Blot, Incubation, Irradiation, Control

Journal: Nitric oxide : biology and chemistry

Article Title: NITRIC OXIDE-MEDIATED RESISTANCE TO PHOTODYNAMIC THERAPY IN A HUMAN BREAST TUMOR XENOGRAFT MODEL: IMPROVED OUTCOME WITH NOS2 INHIBITORS

doi: 10.1016/j.niox.2016.12.003

Figure Lengend Snippet: iNOS/NO-induced resistance of MDA-MB-231 cells to photosensitized toxicity. ALA-treated cells were either not irradiated or irradiated in the absence or presence of 25 μM 1400W (W), using the indicated light fluences. Non-ALA-treated cells were also irradiated as controls. Following treatment, cells were switched to 10% FBS-medium after removal of any detached cells. (A) MTT-assessed viable fraction after 20 h of dark incubation; (□) W absent, (△) W present, (○) light-only control. (B) AnnexinV-FITC-assessed apoptosis after 4 h of dark incubation; 25 μM camptothecin (CPT) served as a reference for induction of complete apoptosis. Data in panels (A) and (B) are means ± SEM of values from three separate experiments. *P<0.02 compared with ALA/hν.

Article Snippet: The polyclonal antibody against human iNOS was obtained from Cayman Chemicals (Ann Arbor, MI).

Techniques: Irradiation, Incubation, Control

Journal: Nitric oxide : biology and chemistry

Article Title: NITRIC OXIDE-MEDIATED RESISTANCE TO PHOTODYNAMIC THERAPY IN A HUMAN BREAST TUMOR XENOGRAFT MODEL: IMPROVED OUTCOME WITH NOS2 INHIBITORS

doi: 10.1016/j.niox.2016.12.003

Figure Lengend Snippet: Accelerated proliferation and invasion of MDA-MB-231 cells surviving an ALA/light challenge: role of iNOS/NO. ALA-treated cells were exposed to a 1 J/cm2 light fluence in the absence vs. presence of 25 μM 1400W or 25 μM cPTIO. Cells treated with ALA alone or 1400W alone served as controls. Twenty-four hours after treatment, cells were switched to fresh FBS-medium. (A) Proliferation assessed by MTT-detectable viable cell number. (B) Invasion assessed by trans-well assay. Cells traversing the trans-well filter and collecting on its lower surface were photographed (upper images), then removed by centrifugation and quantified by MTT assay (lower image). Plotted data in panels (A) and (B) are means ± SEM of values from three replicate experiments. *P<0.01 compared with ALA/hν.

Article Snippet: The polyclonal antibody against human iNOS was obtained from Cayman Chemicals (Ann Arbor, MI).

Techniques: Centrifugation, MTT Assay

Journal: Nitric oxide : biology and chemistry

Article Title: NITRIC OXIDE-MEDIATED RESISTANCE TO PHOTODYNAMIC THERAPY IN A HUMAN BREAST TUMOR XENOGRAFT MODEL: IMPROVED OUTCOME WITH NOS2 INHIBITORS

doi: 10.1016/j.niox.2016.12.003

Figure Lengend Snippet: Altered expression or activity of effector proteins in ALA/light-treated cells: effects of iNOS inhibition. After pre-incubation with ALA, MDA-MB-231 cells were exposed to a 1 J/cm2 light fluence in the absence (ALA/hν) or presence or 25 μM GW274150 (ALA/GW/ hν). ALA-only dark controls (DC) were run alongside. At the indicated post-irradiation times, cells were retrieved for Western analysis of Bcl-xL, Bax, Survivin, S100A4, phosphorylation-activated Akt (p-Akt), total Akt, and β-actin. Number below each effector protein band represents band intensity relative to β-actin and normalized to DC. Total protein load: 50 μg per lane.

Article Snippet: The polyclonal antibody against human iNOS was obtained from Cayman Chemicals (Ann Arbor, MI).

Techniques: Expressing, Activity Assay, Inhibition, Incubation, Irradiation, Western Blot, Phospho-proteomics

Journal: Nitric oxide : biology and chemistry

Article Title: NITRIC OXIDE-MEDIATED RESISTANCE TO PHOTODYNAMIC THERAPY IN A HUMAN BREAST TUMOR XENOGRAFT MODEL: IMPROVED OUTCOME WITH NOS2 INHIBITORS

doi: 10.1016/j.niox.2016.12.003

Figure Lengend Snippet: ALA-PDT suppression of mouse-borne tumor xenografts: improved response after multiple administrations of iNOS inhibitor 1400W (W) or GW274150 (GW). (A) Effects of iNOS inhibition on light-only controls. Female SCID mice bearing MDA-MB-231 tumors were injected i.p. with PBS vehicle or 1400W (10 mg/kg) in PBS. After 4 h, each animal was mildly anesthetized with isoflurane and placed into an opaque restraining cone with a nose opening and a cutout for tumor exposure to irradiation. Omnilux-revive™ irradiation was carried out for 15 min, corresponding to a delivered light fluence of ~95 J/cm2. After irradiation, animals were re-injected with 1400W every day until termination, tumor volumes being measured every third day. The mean ± SEM of values for 3 animals is plotted for each time point. (B) Effects of iNOS inhibition on ALA-PDT. MDA-MB-231 tumor-bearing SCID mice were injected i.p. with ALA (100 mg/kg) or PBS vehicle (light-only controls), followed by 1400W (10 mg/kg) or GW274150 (25 mg/kg). After 4 h in the dark, each animal was mildly anesthetized, placed in a restraining cone, and irradiated as described in (A). After PDT, animals were caged under subdued room light and re-injected with 1400W or GW274150 every day and tumor volumes were measured every third day. For each time point, the mean ± SEM of values for the following animal numbers are plotted: 6 (hν); 6 (ALA/hν); 3 (ALA/W/hν); 3 (ALA/GW/hν). P<0.05 for ALA/W/hν vs. ALA/hν at 6, 9, and 12 days; P<0.05 for ALA/GW/hν vs. ALA/hν at 9 and 12 days.

Article Snippet: The polyclonal antibody against human iNOS was obtained from Cayman Chemicals (Ann Arbor, MI).

Techniques: Inhibition, Injection, Irradiation

Journal: Nitric oxide : biology and chemistry

Article Title: NITRIC OXIDE-MEDIATED RESISTANCE TO PHOTODYNAMIC THERAPY IN A HUMAN BREAST TUMOR XENOGRAFT MODEL: IMPROVED OUTCOME WITH NOS2 INHIBITORS

doi: 10.1016/j.niox.2016.12.003

Figure Lengend Snippet: iNOS upregulation in MDA-MB-231 tumor tissue after ALA-PDT. Tumor-bearing SCID mice were irradiated after prior administration of ALA in PBS (ALA/ hν) or PBS alone (hν, a light-only control). After irradiation as described in Fig. 6, tumors were either harvested immediately (day-0) or after a 2 or 6 days delay, during which animals were kept away from any bright room lighting. Tumors were homogenized, homogenates centrifuged, and supernatants recovered for Western blot analysis of iNOS as described in Sect. 2.9. Each lane in the blots shown represents material from pooled homogenates of duplicate experimental tumors. Numbers below iNOS bands indicate band intensities relative to β-actin and normalized to day zero. Numbers below β-actin bands indicate average size of tumors used for preparing these Western blots. Total tumor protein applied per lane: 40 μg.

Article Snippet: The polyclonal antibody against human iNOS was obtained from Cayman Chemicals (Ann Arbor, MI).

Techniques: Irradiation, Control, Western Blot

Journal: Nitric oxide : biology and chemistry

Article Title: NITRIC OXIDE-MEDIATED RESISTANCE TO PHOTODYNAMIC THERAPY IN A HUMAN BREAST TUMOR XENOGRAFT MODEL: IMPROVED OUTCOME WITH NOS2 INHIBITORS

doi: 10.1016/j.niox.2016.12.003

Figure Lengend Snippet: Effects of ALA-PDT on expression of key pro- vs. anti-survival effectors in tumor samples after ALA-PDT: effects of an iNOS inhibitor. At the indicated times (days) after subjecting MDA-MB-231 tumor-bearing SCID mice to irradiation alone (hν), irradiation after ALA administration (ALA/hν), or irradiation after ALA and GW274150 administration (ALA/GW/hν), tumors were isolated and homogenates prepared. Samples were used for Western blot analysis of Bcl-xL, Bax, Survivin, S100A4, and β-actin levels. Numbers below effector protein bands indicate band intensity relative to β-actin and normalized to day zero, i.e. immediately after PDT. Total tumor protein loaded: 40 μg per lane.

Article Snippet: The polyclonal antibody against human iNOS was obtained from Cayman Chemicals (Ann Arbor, MI).

Techniques: Expressing, Irradiation, Isolation, Western Blot