il 6 protein concentrations  (Boster Bio)

Journal: The FASEB Journal

Article Title: IL-6/p-BTK/p-ERK signaling mediates calcium phosphate–induced pruritus

doi: 10.1096/fj.201900016RR

Figure Lengend Snippet: Scratching and expression of IL-6 and p-ERK in mice injected with CaP. A) The number of hind paw scratches was counted for 2 h after intradermal injection of CaP (0, 0.5 and 5 mg/ml). B) The level of IL-6 in the dorsal skin of ICR mice injected with or without CaP was measured with ELISA. C) Western blot analysis of total ERK (t-ERK 1/2), p-ERK 1/2 and GAPDH in DRG of mice injected with/without CaP. D, E) The ratios of intensities of p-ERK 1/2 (D) and t-ERK 1/2 (E) relative to GAPDH in Western blot analysis were shown. The mean ± se for 3 independent experiments with 4 mice per group was calculated. Ns, not significant. *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: Human sera were collected from supernatants after centrifuging blood at 2800 g for 20 min. IL-6 concentration was determined using a mouse or human IL-6 ELISA assay kit (R&D Systems, Minneapolis, MN, USA) following the manufacturer’s instructions.

Techniques: Expressing, Injection, Enzyme-linked Immunosorbent Assay, Western Blot

Journal: The FASEB Journal

Article Title: IL-6/p-BTK/p-ERK signaling mediates calcium phosphate–induced pruritus

doi: 10.1096/fj.201900016RR

Figure Lengend Snippet: The essential role of IL-6 in the CaP-induced elevation of p-ERK in DRG. A) Bar graphs show the number of scratches postinjection of CaP (0 or 5 mg/ml) in WT and IL-6 KO mice. B) Western blot analyses of p-ERK 1/2 and GAPDH in DRG of WT and IL-6 KO mice. C, D) The ratios of intensities of p-ERK 1/2 (C) and t-ERK 1/2 (D) relative to GAPDH were illustrated. Four mice per group were used for this experiment. The mean ± se for 3 separate experiments was calculated. *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: Human sera were collected from supernatants after centrifuging blood at 2800 g for 20 min. IL-6 concentration was determined using a mouse or human IL-6 ELISA assay kit (R&D Systems, Minneapolis, MN, USA) following the manufacturer’s instructions.

Techniques: Western Blot

Journal: The FASEB Journal

Article Title: IL-6/p-BTK/p-ERK signaling mediates calcium phosphate–induced pruritus

doi: 10.1096/fj.201900016RR

Figure Lengend Snippet: Suppression of the CaP-induced scratching, IL-6 elevation, and p-ERK activation by BTK inhibition. A–C) The number of scratches (A), the levels of IL-6 (B), and protein expressions of p-ERK 1/2, t-ERK ½, and GAPDH in Western blot analysis (C) in mice injected with CaP (5 mg/ml) in the presence or absence of ibrutinib (Ib), a BTK inhibitor, were demonstrated. D, E) The ratios of intensities of p-ERK 1/2 (D) and t-ERK 1/2 (E) relative to GAPDH were illustrated. Four mice per group were used for this experiment. The mean ± se from 3 separate experiments was calculated. *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: Human sera were collected from supernatants after centrifuging blood at 2800 g for 20 min. IL-6 concentration was determined using a mouse or human IL-6 ELISA assay kit (R&D Systems, Minneapolis, MN, USA) following the manufacturer’s instructions.

Techniques: Activation Assay, Inhibition, Western Blot, Injection

Journal: The FASEB Journal

Article Title: IL-6/p-BTK/p-ERK signaling mediates calcium phosphate–induced pruritus

doi: 10.1096/fj.201900016RR

Figure Lengend Snippet: Skin lesions in mouse skin injected with CaP and morphologies and IL-6 expression in the skin of healthy subjects and the nonitchy or itchy skin of patients with CKD. A) The skin morphologies of ICR mice after injection with H2O and CaP (5 mg/ml) for 7 d were displayed. Skin lesions were indicated by red arrows. Scale bar, 5 mm. B) Skin images of the back of the hand from healthy subjects (normal) and the nonitchy or itchy skin of patients with CKD were shown. Xerotic skin with white spots were pointed out with red arrows. Scale bar, 1 mm. C) Immunostaining of IL-6 (green dots and red arrows) in the skin of healthy subjects (normal) and in the nonitchy or itchy skin of patients with CKD. Nuclei were labeled with DAPI (blue dots). Dotted lines indicate the epidermal layers. Scale bars, 100 μm. D) Quantification of green dots in a selected area. The mean ± se was determined using at least 3 skin biopsies. *P < 0.05, ***P < 0.001.

Article Snippet: Human sera were collected from supernatants after centrifuging blood at 2800 g for 20 min. IL-6 concentration was determined using a mouse or human IL-6 ELISA assay kit (R&D Systems, Minneapolis, MN, USA) following the manufacturer’s instructions.

Techniques: Injection, Expressing, Immunostaining, Labeling

Journal: The FASEB Journal

Article Title: IL-6/p-BTK/p-ERK signaling mediates calcium phosphate–induced pruritus

doi: 10.1096/fj.201900016RR

Figure Lengend Snippet: Activation of p-BTK and p-ERK by IL-6. A) Western blot analysis of protein expressions of p-BTK, BTK, p-ERK 1/2, t-ERK 1/2, and GAPDH in DRG cells treated with mouse recombinant IL-6 (100 ng/ml) for 0, 0.5, 1, 5, 10, and 30 min. B, C) The ratios of intensities of p-BTK (B) and p-ERK 1/2 (C) relative to GAPDH were illustrated. D, E) Immunostaining (D) of p-BTK (green dots; arrows) and nuclei (DAPI blue dots) and quantification (E) of fluorescent intensities of p-BTK in DRG cells treated with or without IL-6 for 1 min. Scale bars, 100 μm. F) Western blot analysis of the expressions of p-BTK, BTK, p-ERK 1/2, t-ERK 1/2, and β-actin in DRG cells treated with or without IL-6 in the presence or absence of ibrutinib (Ib). G, H) The ratios of intensities of p-BTK (G) and p-ERK 1/2 (H) relative to β-actin were displayed. The mean ± se from 3 separate experiments was calculated (ns, not significant). ***P < 0.001.

Article Snippet: Human sera were collected from supernatants after centrifuging blood at 2800 g for 20 min. IL-6 concentration was determined using a mouse or human IL-6 ELISA assay kit (R&D Systems, Minneapolis, MN, USA) following the manufacturer’s instructions.

Techniques: Activation Assay, Western Blot, Recombinant, Immunostaining

Journal: The FASEB Journal

Article Title: IL-6/p-BTK/p-ERK signaling mediates calcium phosphate–induced pruritus

doi: 10.1096/fj.201900016RR

Figure Lengend Snippet: The signaling pathway of CaP-induced scratching, elevation of IL-6, activation of p-BTK and p-ERK, and a possible positive feedback loop between IL-6 and p-BTK is proposed.

Article Snippet: Human sera were collected from supernatants after centrifuging blood at 2800 g for 20 min. IL-6 concentration was determined using a mouse or human IL-6 ELISA assay kit (R&D Systems, Minneapolis, MN, USA) following the manufacturer’s instructions.

Techniques: Activation Assay