Journal: Scientific reports

Article Title: The N-Acetyl Phenylalanine Glucosamine Derivative Attenuates the Inflammatory/Catabolic Environment in a Chondrocyte-Synoviocyte Co-Culture System.

doi: 10.1038/s41598-019-49188-9

Figure Lengend Snippet: Figure 5. IL-1β increases IKKα nuclear translocation and phosphorylation of serine 10 on histone H3, while NAPA attenuates these signalling events. (A) Overlapping signals of nuclear counterstaining (DAPI) and IKKα detected via an Alexa Fluor 555 secondary antibody: the colocalized signals indicate that NAPA addition is effective in reducing nuclear translocation of IKKα. (B) Upper pictures: left, western blot of anti- phosphorylated serine 10 of histone H3, along with GAPDH as a loading control and right: densitometric analysis of the signal showing the different pattern of H3pSer10 accumulation in CTR (circle), IL-1β (square) or IL-1β + NAPA (triangle) conditions. Lower images: specificity of the signal obtained with the anti-H3 phosphorylated serine 10: 20x field pictures of chondrocytes grown on coverslips in the bottom of wells at time 0 in control (upper row) or IL-1β stimulated conditions (lower row): Green: IKKα detected with an Alexa Fluor 488 anti-rabbit antibody; red: H3pSer10 signal detected with an Alexa Fluor 555 anti-mouse antibody; blue: nuclear DNA stained with Hoechst 33342 and merged images.

Article Snippet: IKKα staining was performed with 5 μg/ml rabbit anti-IKKα antibody (BOSTER Biological Technology, Freemont, CA, USA), while the extent of phosphorylation of serine 10 of histone H3 was evaluated with 5 μg/ml anti-phospho-histone H3, (Ser10), (mouse monoclonal, Upstate–Millipore) overnight at 4 °C.

Techniques: Translocation Assay, Phospho-proteomics, Western Blot, Control, Staining

Journal: PeerJ

Article Title: Foxa2 attenuates steatosis and inhibits the NF-κB/IKK signaling pathway in nonalcoholic fatty liver disease

doi: 10.7717/peerj.16466

Figure Lengend Snippet: (A) Protein expression of Foxa2. (B) Protein expression of p-NF-κB, NF-κB, p-IKK, and IKK. (C) Protein expression of FAS, ACC, and CPT1α were detected by western blotting. HepG2 cells were treated with OA, oe-Foxa2, and lipopolysaccharide (LPS, 1 µg/mL). ** P < 0.01 compared with Control; ## P < 0.01 compared with OA. & P < 0.05 and && P < 0.01 compared with OA + oe-Foxa2 + LPS.

Article Snippet: The primary antibodies including carnitine palmitoyl transferase 1α (CPT1α; 1:1000; DF12004), Foxa2 (1:1000; DF13363), NF-κB (1:2000; AF5006), p-NF-κB (1:2000; AF2006), IKK (1:2000; AF6014), and p-IKK (1:2000; AF3013) were purchased from Affinity Biosciences (JIangsu, China); acetyl-CoA carboxylase (ACC; 1:1000; 3676) and GAPDH (1:1000; 2118) were purchased from Cell Signaling Technology (Waltham, MA, USA); fatty acid synthase (FAS; 1:1000; E-AB-40063) was purchased from Elabscience Biotechnology Co., Ltd (Wuhan, China).

Techniques: Expressing, Western Blot, Control