Journal: Molecules (Basel, Switzerland)

Article Title: Hispidulin Inhibits the Vascular Inflammation Triggered by Porphyromonas gingivalis Lipopolysaccharide.

doi: 10.3390/molecules28186717

Figure Lengend Snippet: Figure 2. Hispidulin reduces ICAM-1 expression in P. gingivalis LPS-induced HUVECs. (a,b) HUVECs were exposed to P. gingivalis LPS (5 µg/mL) alone, hispidulin alone (5 µM), or a combination of P. gingivalis LPS and hispidulin for 16 h, and the expression of ICAM-1 was analyzed using RT-qPCR and Western blotting. The control level was established at 1.0, and the measurements were adjusted relative to β-actin for normalization (a) or α-tubulin as the internal control. The quantification of the ICAM-1 protein level was obtained with densitometry and normalized to α-tubulin (b). * p < 0.01 compared to that of control. # p < 0.01 compared to that of P. gingivalis LPS. Data shown are the mean ± SD, obtained from at least three independent experiments. (c) HUVECs were exposed to P. gingivalis LPS (5 µg/mL) alone, hispidulin alone (5 µM), or a combination of P. gingivalis LPS and hispidulin for 16 h. ICAM-1 (CD54) was quantified using flow cytometry.

Article Snippet: Antibodies against human ICAM-1 and α-tubulin were obtained from Santa Cruz Biotechnology (#SC-8439, Dallas, TX, USA) and Bioworld Technology (#BS1699, St. Louis Park, MN, USA), respectively.

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Control, Cytometry

Journal: Molecules (Basel, Switzerland)

Article Title: Hispidulin Inhibits the Vascular Inflammation Triggered by Porphyromonas gingivalis Lipopolysaccharide.

doi: 10.3390/molecules28186717

Figure Lengend Snippet: Figure 4. Hispidulin inhibits P. gingivalis LPS-induced NF-κB-dependent ICAM-1 promoter activity, MAPKs, and AKT signaling. (a) HUVECs were transiently transfected with the full-length and truncated promoters of the ICAM-1 gene. Transfected HUVECs were exposed to P. gingivalis LPS (5 µg/mL) alone, hispidulin (5 µM) alone, or a combination of P. gingivalis LPS and hispidulin for 16 h. * p < 0.01 compared to that of the control. # p < 0.01 compared to that of P. gingivalis LPS. Data shown are the mean ± SD, obtained from at least three independent experiments. (b) HUVECs were pretreated with hispidulin (5 µM) for 30 min and then incubated for 1 h with P. gingivalis LPS (5 µg/mL). NF-κB p65 (green) localization in the nuclei (blue) was observed. (c) HUVECs were pretreated for 30 min with or without hispidulin (5 µM) before stimulation with P. gingivalis LPS (5 µg/mL) for 10 min. Anti-phospho-ERK, anti-ERK, anti-phospho-JNK, anti-JNK, anti-phospho-p38MAPK, anti-p38MAPK, anti-phospho-AKT, and anti-AKT antibodies were used to probe the Western blots. α-tubulin served as the internal control.

Article Snippet: Antibodies against human ICAM-1 and α-tubulin were obtained from Santa Cruz Biotechnology (#SC-8439, Dallas, TX, USA) and Bioworld Technology (#BS1699, St. Louis Park, MN, USA), respectively.

Techniques: Activity Assay, Transfection, Control, Incubation, Western Blot

Journal: International Journal of Biological Sciences

Article Title: Selective targeting of the TLR4 co-receptor, MD2, prevents colon cancer growth and lung metastasis

doi: 10.7150/ijbs.39098

Figure Lengend Snippet: MD2 blockade inhibits NF-κB activation in AOM/DSS mouse model. WT or MD2 -/- mice were treated with AOM/DSS to produce a colitis-associated colon acncer growth (Methods). WT mice were also orally administered L6H21 (60 mg/ml) or 1% CMC-Na (vehicle control); n=10. (A) Western blot analysis of phosphorylated IκB-α and phosphorylated NF-κB P65 subunit (p-P65) in mouse colon tissues; GAPDH used as loading control. The immune-reactive bands were quantified by densitometry using Image J analysis: (B) p-IκB-α and (C) p-P65; values normalized to GAPDH. (D) Immunohistochemical staining of colon tissues from mice for MD2 (upper row; brown) and phosphorylated p65 subunit of NF-κB (bottom row; brown); n=6. Quantification (data in D) of MD2 intensity and phosphorylated NF-κB p65 subunit are provided in supplementary file. Real-time qPCR determination of pro-inflammatory genes in colonic tissue; mRNA values normalized to β-actin and reported relative to WT Con: (E) IL-6, (F) TNF-α, (G) TGF-β; n=3. (H) ELISA detection of IL-6 in colon tissue lysates; values normalized to total protein; n=6. Real-time qPCR determination of adhesion molecules in colonic tissue; mRNA values normalized to β-actin and reported relative to WT Con: (I) ICAM-1, (J) VCAM-1; n=3. Data in B, C, E, F, G, H, I, J are shown as mean±SEM; * P <0.05, ** P <0.01, *** P <0.001.

Article Snippet: The primers for IL-6, TNF-α, TGF-β, ICAM-1, VCAM-1 and β-actin were obtained from Invitrogen, and are shown in Supplementary file ( ).

Techniques: Activation Assay, Western Blot, Immunohistochemical staining, Staining, Enzyme-linked Immunosorbent Assay