Journal: International journal of molecular sciences

Article Title: WGA-M001, a Mixture of Total Extracts of Tagetes erecta and Ocimum basilicum , Synergistically Alleviates Cartilage Destruction by Inhibiting ERK and NF-κB Signaling.

doi: 10.3390/ijms242417459

Figure Lengend Snippet: Figure 4. WGA-M001 regulated OA-related molecules through dephosphorylation in the NF-κB and ERK pathways. For in silico analysis, 200 µg/mL of Tagetes erecta, Ocimum basilicum, or WGA-M001 was used for treatment of chondrocytes for 12 h along with IL-1β (1 ng/mL), and RNA sequencing was performed. (A) In each signaling pathway, the number of genes upregulated by IL-1β and then downregulated by T. erecta, O. basilicum, or WGA-M001 was shown. (B,C) Protein levels of pp38, p38, pJNK, JNK, pERK, ERK, pp65, p65, and IκB were detected by Western blot analysis and relative intensities were quantified by densitometry (n = 5). Each protein level was normalized to ERK. Data are represented as mean ± SD as results of analysis by one-way ANOVA with Dunnett’s multiple comparisons test (n = 5). * p < 0.05, **** p < 0.0001, and n.s = not significant.

Article Snippet: 2023, 24, 17459 12 of 16 cam), rabbit anti-COX-2 (ab52237; Abcam), mouse anti-IκB (9242; Cell Signaling Technology (CST), Danvers, MA, USA), mouse anti-p65 (#6956; CST), mouse anti-pp65 (#13346; CST), mouse anti-p38 (#9212; CST), mouse anti-pp38 (#9215S; CST), mouse anti-c-JNK (#9252S; CST), mouse anti-pJNK (#9251S; CST), and mouse anti-pERK (#9101S; CST).

Techniques: De-Phosphorylation Assay, In Silico, RNA Sequencing, Western Blot

Journal: Mediators of Inflammation

Article Title: MCL Plays an Anti-Inflammatory Role in Mycobacterium tuberculosis -Induced Immune Response by Inhibiting NF- κ B and NLRP3 Inflammasome Activation

doi: 10.1155/2017/2432904

Figure Lengend Snippet: MCL inhibits Mtb-triggered NF- κ B activation. Raw264.7 cells (1 × 10 6 /well) were seeded in 6-well plates and infected with Mtb for 4 hrs. Cells were washed by PBS and treated with MCL (10 μ M) for indicated time courses. Phosphorylation of p65 and β -actin was detected by Western blot (a). The lower bar graph showed the statistical results for phosphorylation levels of NF- κ B p65 (a). Data are presented as mean ± SD in at least three independent experiments, ∗∗ p < 0.01. Raw264.7 cells (3 × 10 5 /well) were cotransfected with NF- κ B luciferase reporter plasmid and pRL-TKR Renilla luciferase plasmid as previous study . After 30 hr, cells were infected with H37Ra at MOI of 10 and treated with MCL (10 μ M) for 12 hr, and the luciferase activities were measured (b). Data are shown with the means ± SD ( n ≧3). ∗ p < 0.05. Raw264.7cells (3 × 10 5 /well) were plated in climbing slice and followed by appropriate treatment. Immunofluorescence assay was conducted with rabbit anti-pp65 antibody (green) and DAPI (blue) (c). The results represented three independent experiments.

Article Snippet: Rabbit anti-ASC (10500-1-AP, proteintech), anti-LC3 (sc-16755, Santa Cruz Biotechnology), anti-caspase-1 (sc-514, Santa Cruz Biotechnology), and anti-pp65 (cst-3033, cell signaling technology) antibodies were used for immunoblot.

Techniques: Activation Assay, Infection, Western Blot, Luciferase, Plasmid Preparation, Immunofluorescence

Journal:

Article Title: Human cytomegalovirus gene expression during infection of primary hematopoietic progenitor cells: A model for latency

doi: 10.1073/pnas.252630899

Figure Lengend Snippet: GFP is a reliable marker for TLsubUL21.5 infection of hematopoietic cells. Mononuclear cells enriched for CD34+ cells (≈60%) were mock-infected (A) or infected with TLsubUL21.5 at a multiplicity of 5 plaque-forming units (pfu) per cell (B). At 10 h postinfection, cells were analyzed for expression of GFP and staining for pp65 by flow cytometry. Data from a representative experiment are shown.

Article Snippet: Intracellular staining for the pp65 virion protein was performed by fixing and permeablizing cells with Fix and Perm (Caltag, South San Francisco, CA), and then reacting with an anti-pp65 mouse monoclonal antibody followed by a goat anti-mouse secondary IgG conjugated to allophycocyanin (APC; BD Biosciences).

Techniques: Marker, Infection, Expressing, Staining, Flow Cytometry