Journal: Molecular medicine reports

Article Title: Mechanisms of N‑acetylcysteine in reducing monocrotaline‑induced pulmonary hypertension in rats: Inhibiting the expression of Nox1 in pulmonary vascular smooth muscle cells.

doi: 10.3892/mmr.2017.7326

Figure Lengend Snippet: Figure 2. Expression of Nox 1 and Nox4. (A) The mRNA level of Nox1 and Nox4 in lung tissue. The standardization of group Cont was 1. #P<0.05 vs. Cont; *P<0.05 vs. MCT; **P<0.001 vs. MCT. (B) The mRNA level of Nox1 and Nox4 in PASMCs. The standardization of group Cont was 1. $P<0.05 vs. C1; +P<0.001 vs. M1. (C) Double immunofluorescence staining of Nox1 and α‑SMA in lung tissue. Green, α‑SMA; red, Nox1. (D) Western blotting and (E) densi tometric analysis of the expression of Nox1 protein in PASMCs (n=6). β‑actin was used as the internal control. $$P<0.001 vs. C1; ++P=0.011 vs. M1. (F) DHE fluorescence intensity of the lung tissue. (G) The standardization of group Cont was 1. The data demonstrate the fold change of group MCT and group NAC relative to group Cont. ##P<0.001 vs. Cont; **P<0.001 vs. MCT. (H) Correlation analysis of Nox1 and DHE fluorescence intensity. r=0.850; P<0.001. PASMC, pulmonary artery smooth muscle cell; Nox, NADPH oxidase; Cont, control; α‑SMA, smooth muscle actin; MCT, monocrotaline; NAC, N‑acetylcysteine; M, MCT‑treated; N, NAC‑treated group; C, control subgroup; DHE, dihydroethidium.

Article Snippet: Excess serum was absorbed and primary anti-Nox1 antibody (BA3720; 1:100; Boster Biological Technology) added for overnight incubation at 4 ̊C.

Techniques: Expressing, Double Immunofluorescence Staining, Western Blot, Control, Fluorescence

Journal: Molecular Medicine Reports

Article Title: Heme oxygenase 1‑overexpressing bone marrow mesenchymal stem cell‑derived exosomes suppress interleukin‑1 beta‑induced apoptosis and aging of nucleus pulposus cells

doi: 10.3892/mmr.2025.13481

Figure Lengend Snippet: Activation of the NF-κB signalling in NP cells induced by IL-1β was repressed after Exo-HO-1 treatment. Western blotting was performed to detect p-p65 and/or p65 protein levels in NP cells with different treatments (control, IL-1β, IL-1β + Exo and IL-1β + Exo-HO-1) as well as in the nuclear (N) and cytoplasmic (C) fractions of the cells. Histone H3 was used as an internal reference for the nucleus. ***P<0.001 vs. control; # P<0.05, ## P<0.01 and ### P<0.001 vs. IL-1β. NP, nucleus pulposus; IL, interleukin; Exo, exosomes; HO-1, heme oxygenase 1; p-phosphorylated.

Article Snippet: After blocking with a rapid blocking solution (Beyotime Institute of Biotechnology) at 4°C overnight, the membranes were incubated with appropriate primary antibodies including anti-HO-1 (cat. no. GTX637432; 1:2,000; GeneTex, Inc.), anti-TGS101 (cat. no. A01233-2, 0.25 μg/ml; Boster Bio), anti-CD63 (FNab01490; 1:1,000; Fine Biotech Co., Ltd, Wuhan, China) and anti-Cainexin (cat. no. GTX109669; 1:5,000; GeneTex, Inc.), Bcl-2 (cat. no. FNab00839; 1:1,000; Wuhan Fine Biotech Co., Ltd.), Bax (cat. no. FNab00810; 1:2,000; Wuhan Fine Biotech Co., Ltd.), cleaved caspase 3 (cat. no. MBS9410752; 1:1,000; MyBioSource, Inc.), anti-p65 (cat. no. PB9324; 0.5 μg/ml; Boster Bio), anti-p-p65 (cat. no. A00284S468-2; 1:2,000; Boster Bio), anti-GAPDH (cat. no. H00227; 1:5,000; Boster Bio) and anti-Histone H3 (cat. no. M12477-9; 1:1,000; Boster Bio) antibodies.

Techniques: Activation Assay, Western Blot, Control

Journal: PLoS ONE

Article Title: Deficiency of NOX1 or NOX4 Prevents Liver Inflammation and Fibrosis in Mice through Inhibition of Hepatic Stellate Cell Activation

doi: 10.1371/journal.pone.0129743

Figure Lengend Snippet: Primer Sequences for Real-time.

Article Snippet: The NOX1 and NOX4 antibodies were purchased from Boster Biological Technology Company, Wuhan, China.

Techniques:

Journal: PLoS ONE

Article Title: Deficiency of NOX1 or NOX4 Prevents Liver Inflammation and Fibrosis in Mice through Inhibition of Hepatic Stellate Cell Activation

doi: 10.1371/journal.pone.0129743

Figure Lengend Snippet: (A) HSC proliferation is reduced in NOX1KO and NOX4KO mice after CCl 4 injury. HSC proliferation in liver was presented by PCNA immunofluorescence microscopy. Desmin is a specific marker of HSCs (Red). PCNA is a marker of proliferation (Green). Original magnification X10. (B) Proliferation was reduced in HSCs lacking NOX1 and NOX4 in response to PDGF (10 ng/ml) for 24 h. Proliferative HSCs were presented by Ki67 (Red) immunofluorescence microscopy. Nuclei were presented by DAPI (Blue). Original magnification X10. (C) Graph of PCNA-positive HSCs in vivo. (D) Graph of Ki67-positive HSCs in vitro.

Article Snippet: The NOX1 and NOX4 antibodies were purchased from Boster Biological Technology Company, Wuhan, China.

Techniques: Immunofluorescence, Microscopy, Marker, In Vivo, In Vitro

Journal: PLoS ONE

Article Title: Deficiency of NOX1 or NOX4 Prevents Liver Inflammation and Fibrosis in Mice through Inhibition of Hepatic Stellate Cell Activation

doi: 10.1371/journal.pone.0129743

Figure Lengend Snippet: Human livers from controls (N = 7) and patients with cirrhosis (N = 10) were analyzed by Sirius Red staining and by immunofluorescence with antibodies against NOX1 and NOX4. The graphs show the percent positive area of the staining or immunofluorescence expressed as mean+S.D., p<0.05 for each comparison of control vs. fibrosis.

Article Snippet: The NOX1 and NOX4 antibodies were purchased from Boster Biological Technology Company, Wuhan, China.

Techniques: Staining, Immunofluorescence, Comparison, Control

Journal: Scientific Reports

Article Title: Promotion of the inflammatory response in mid colon of complement component 3 knockout mice

doi: 10.1038/s41598-022-05708-8

Figure Lengend Snippet: Expression levels of members in the NF-κB signaling pathway. ( A ) Expression levels of NF-κB-p65 and IκB-α proteins were determined by Western blot analysis using specific primary antibody and HRP-labeled anti-rabbit IgG antibody. ( B ) Band intensities were determined using an imaging densitometer, and protein expressions were calculated relative to the intensity of β-actin. Tissue samples were collected from 3 to 5 mice per group, and each lysate was analyzed in duplicate for Western blot (final n = 6–10). Data are reported as the mean ± SD. * indicates p < 0.05 compared to the WT mice.

Article Snippet: Proteins (30 μg) were then separated by 4–20% sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE) for 2 h, and subsequently transferred to nitrocellulose membranes for 3 h at 40 V. Each membrane was then incubated separately, overnight at 4 °C, with the following primary antibodies: anti-C3 (ab200999, Abcam Com.), anti-C3aR (bs-2955R, Bioss Inc.), anti-CR1 (LS-C777464, LSBio Inc.), anti-COX-2 (12282, Cell Signaling Technology Inc., Danvers, MA, USA), anti-NLRP3 (15101, Cell Signaling Technology Inc.), anti-Cas 1 (24232, Cell Signaling Technology Inc.), anti-ASC (67824, Cell Signaling Technology Inc.), anti-iNOS (PA3-0304, Thermo Fisher Scientific Inc.), anti-ERK1/2 (9102, Cell Signaling Technology Inc.), anti-p-ERK (E-4) (9101, Santa Cruz Biotechnology Inc., Dallas, TX, USA), anti-JNK (9252, Cell Signaling Technology Inc.), anti-p-JNK (9251, Cell Signaling Technology Inc.), anti-p38 (9212, Cell Signaling Technology Inc.), anti-p-p38 (9211, Cell Signaling Technology Inc.), anti-p-NF-κB-p65 (A00284T254, Boster Bio Inc., Pleasanton, CA, USA), anti-IκB (9242, Cell Signaling Technology Inc.), anti-p-IκB (9246S, Cell Signaling Technology Inc.), anti-IL-6 (SC-1265, Santa Cruz Biotechnology Inc.), anti-C5 (ab11898, Abcam Com.), anti-thrombin (ab92621, Abcam Com.), anti-E-cadherin (24E10, Cell Signaling Technology Inc.) or anti-β-actin (4967, Sigma-Aldrich Co.).

Techniques: Expressing, Western Blot, Labeling, Imaging

Journal: Acta biochimica et biophysica Sinica

Article Title: Therapeutic effect of artemisinin on lupus nephritis mice and its mechanisms.

doi: 10.1093/abbs/gmq101

Figure Lengend Snippet: Figure 2 Expression of NF-kBp65 protein, NF-kB, and TGF-b1 mRNA in renal tissue (A) Activity of NF-kB. (B) NF-kBp65 protein expression. (C) TGF-b1 mRNA expression. In the three figures, (1) stands for the mice in group (i), (2) for the mice in group (ii), (3) for the mice in group (iii), and (4) for the mice in group (iv).

Article Snippet: The membrane was incubated with mouse anti-NF-kBp65 monoclonal antibody (Wuhan Boster Biological Technology, Ltd., Wuhan, China) (1: 200) and then with HRP-conjugated anti-mouse IgG secondary antibody (Wuhan Boster Biological Technology, Ltd.) (1: 1000).

Techniques: Expressing, Activity Assay

Journal: The Laryngoscope

Article Title: Cyclooxygenase-2 Signaling in Vocal Fold Fibroblasts

doi: 10.1002/lary.21017

Figure Lengend Snippet: Interleukin (IL)-1β treatment for 1 hour increased transcription of the p50 and p65 subunits of nuclear factor (NF)-κB (A) (representative gel) and (B) increased nuclear translocation of both subunits (n = 3; *P < .05) at 1 hour.

Article Snippet: NF-κB Activation and Nuclear Translocation Activated nuclear NF- κ B in HVOX cells 1 hour following treatment with IL-1 β was assayed using commercially available ELISAs for both the p50 and p65 subunits (Active Motif, Carlsbad, CA).

Techniques: Translocation Assay

Journal: The Laryngoscope

Article Title: Cyclooxygenase-2 Signaling in Vocal Fold Fibroblasts

doi: 10.1002/lary.21017

Figure Lengend Snippet: Pretreatment (2 hours) with inhibitors of the p50 and p65 subunits of nuclear factor (NF)-κB decreased interleukin (IL)-1β-induced nuclear translocation of each subunit (A) (n = 3). Inhibition of the p50 and p65 subunits individually had no effect on IL-1β-induced cyclooxygenase-2 (COX-2) mRNA expression. (B) Combined inhibition of both subunits abrogated this effect (representative gel).

Article Snippet: NF-κB Activation and Nuclear Translocation Activated nuclear NF- κ B in HVOX cells 1 hour following treatment with IL-1 β was assayed using commercially available ELISAs for both the p50 and p65 subunits (Active Motif, Carlsbad, CA).

Techniques: Translocation Assay, Inhibition, Expressing

Journal: Clinical and Experimental Immunology

Article Title: Immunomodulatory glc/man‐directed Dolichos lablab lectin (DLL) evokes anti‐tumour response in vivo by counteracting angiogenic gene expressions

doi: 10.1111/cei.12959

Figure Lengend Snippet: Translational down‐regulation of tumoral angiogenic gene expression by Dolichos lablab lectin (DLL). Immunoblots, gelatin zymography and immunohistochemistry (IHC) were carried out using in‐vivo tumour cells of both Dalton's ascites lymphoma (DLA) and Dalton's lymphoma solid tumour (DLS) with or without DLL treatment in two different concentrations. (a,b) Immunoblots showing altered translational expression of proangiogenic genes such as nuclear factor kappa B (NF‐κB), inhibitory kappa B (I‐κB), hypoxia inducible factor 1α (HIF‐1α), vascular endothelial growth factor (VEGF)‐A, matrix metalloproteinase (MMP)‐2 and 9 in ascitic (panel 1) and solid tumour (panel 2). The graphs indicate the densitometric values of corresponding Western blot data. (c) Reduction of secreted serum VEGF levels in DLL‐treated DLA mice in vivo. (d) Gelatin zymography showing reduced gelatinolytic activity in DLL‐treated mice. (e) DLL‐conditioned medium (DLL‐CM) inhibits the migration of A549 cell in vitro. IHV should be replaced with IHC detection of DLL‐induced altered gene expression. (f,i) IHC detection of proangiogenic gene expression in solid tumours in vivo after DLL treatment. The intense brown staining indicates the respective protein expression. Representative graphs demonstrating the gene expression corresponding to the IHC data are given below. (j) Representative counter haematoxylin‐stained section. Data are represented as mean ± standard error of the mean (s.e.m.) of three independent experiments. Statistically significant values are expressed as *P < 0·05 and **P < 0·01. [Colour figure can be viewed at wileyonlinelibrary.com].

Article Snippet: The materials used were lung adenocarcinoma (A549), cervical carcinoma (SiHa, CaSki) and Ehrlich ascites carcinoma (EAC) from the National Centre for Cell Science (NCCS), Pune, India; squamous cell carcinoma (A388) from the National Centre for Biological Science (NCBS), Bengaluru, India and murine Dalton's ascites lymphoma (DLA) cells, a kind gift from Dr Sathees C. Raghavan, Indian Institute of Science (IISc), Bengaluru, India; and human umbilical vascular endothelial cells (HUVEC), endothelial cell growth media (EGM), sodium bicarbonate (NaHCO 3 ), cyanogen bromide (CNBr) activated Sepharose 4B, matrigel [extracellular matrix (ECM) gel], hydron polymer poly‐hydroxyethyl‐methacrylate (poly‐HEMA), Ficoll histopaque, 3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyltetrazoliumbromide (MTT), anti‐vascular endothelial growth factor (VEGF) and anti‐mouse/rabbit immunoglobulin (Ig)G antibodies, protease inhibitor cocktail and Freund's adjuvants were obtained from Sigma‐Aldrich (St Louis, MO, USA), anti‐IL‐2, anti‐hypoxia inducible factor 1α (HIF‐1α); anti‐MMP‐2 and 9 antibodies from Santa Cruz Laboratories (Santa Cruz, CA, USA); anti‐nuclear factor kappa B (NF‐κB), anti‐IκB and anti‐β‐actin from BD Biosciences (San Jose, CA, USA); RPMI medium, antibiotic–anti‐micotic solution, fetal bovine serum (FBS) from Invitrogen (Carlsbad, CA, USA); anti‐CD‐31 antibody and immunostaining kit from Leica Biosystems (Wetzlar, Germany).

Techniques: Expressing, Western Blot, Zymography, Immunohistochemistry, In Vivo, Activity Assay, Migration, In Vitro, Staining