Journal: Poultry Science

Article Title: mRNA and miRNA expression profiles reveal the potential roles of RLRs signaling pathway and mitophagy in duck hepatitis A virus type 1 infection

doi: 10.1016/j.psj.2024.103839

Figure Lengend Snippet: DHAV-1 activated the RLRs signaling pathway and inflammatory response in duckling livers. (A) Western blot analysis was used to reconfirm RLRs signaling pathway including RIG-I, IRF7, and MAVS. (B) The level of MAVS in the mitochondria were determined by western blot assays. (C) Western blotting analysis for IL-6 and TNF-a levels in duckling livers. a–d Values within a column without the same superscripts are significant different ( P < 0.05).

Article Snippet: Primary antibodies included anti-rabbit RIG-1 (1:2,000, 20,566-1-AP, Proteintech), IRF7 (1:2,000, 22,392-1-AP, Proteintech), MAVS (1:2000, 14341-1-AP, Proteintech), TNF-α (1:1,000, WL01581, Wanleibio), p62/SQSTM1 (1:1,000, T55546, Abmart), LC3 (1:1,000, 4,108, Cell Signaling Technology), PINK1 (1:1,000, WL04963, Wanleibio), Parkin (1:1,000, WL02512, Wanleibio), GSDMD (1:1,000, 310,201-T36, Sino Biological), Caspase 1 (1:1,000, WLH4550, Wanleibio), IL-1β (1:500, BS65005, Bioworld), Caspase 3 (1:1000, WL04004, Wanleibio), COX IV (1:1000, WL02203, Wanleibio), β-Actin (1:4,000, AC026, Abclonal), and anti-mouse IL-6 (1:1,000, 10,395-MM19, Sino Biological).

Techniques: Western Blot

Journal: Theranostics

Article Title: Longitudinal Multiplexed Measurement of Quantitative Proteomic Signatures in Mouse Lymphoma Models Using Magneto-Nanosensors

doi: 10.7150/thno.20706

Figure Lengend Snippet: Development of multiplexed magneto-nanosensor cytokine assays. (A) Optical image of an 8 × 8 array of magneto-nanosensor chip. The chip consists of 64 individually-accessible magneto-nanosensors. Inset: optical image of an individual magneto-nanosensor. (B) Schematic of sandwich protein immunoassays using magneto-nanosensors. (1) Different capture antibodies that specifically capture their target proteins were immobilized on different magneto-nanosensors. Bovine serum albumin (BSA) and biotinylated BSA (Biotin, not shown here) were also immobilized on different sensors as negative and positive controls, respectively. (2) Serum containing target proteins was added to the magneto-nanosensor chip, and allowed to bind to the corresponding capture antibodies. (3) After washing unbound proteins, biotinylated detection antibodies were added to the chip, and allowed to bind to the bound proteins to form sandwich complexes. (4) After unbound detection antibodies were washed, the chip was loaded into a reader station, and streptavidin-coated MNPs were introduced. The binding signals of MNPs to the sandwich complexes were monitored and recorded by the reader station. (C) Real-time monitoring of binding signals of MNPs. The baseline signals were measured after the chip was loaded into the reader station. Then, MNPs were introduced to the chip as indicated by the gray arrow. The binding signals were monitored until they reached their plateaus. Six recombinant proteins (GCSF: 1 ng/mL, TNF-α: 1 ng/mL, Eotaxin: 1 ng/mL, FLT3LG: 1 ng/mL, IL-6: 0.1 ng/mL, and VEGF: 0.1 ng/mL) were used as a test sample. The error bars represent standard deviations of 4 identical magneto-nanosensors. (D) Titration curves of 6-plex protein assays measured by multiplexed magneto-nanosensors. Six recombinant proteins were mixed to be at indicated final concentrations (10 and 1 ng/mL, and 100, 10, and 1 pg/mL), and measured using multiplexed magneto-nanosensor protein assays. Each data point is the average signal of 4 identical magneto-nanosensors, and titration curves (solid lines) were calculated using four parameter logistic (4-PL) regression (IL-6: R 2 = 0.9997, GCSF: R 2 = 0.9892, Eotaxin: R 2 = 0.9936, TNF-α: R 2 = 0.9950, FLT3LG: R 2 = 0.9524, and VEGF: R 2 = 0.9969).

Article Snippet: After rinsing the chip again, 50 μL of a cocktail of 6 different biotinylated detection antibodies (anti-IL-6: BAF406, anti-GCSF: BAF414, anti-Eotaxin: BAF420, anti-TNF-α: BAF410, anti-FLT3LG: BAF427, and anti-VEGF: BAF493 from R&D Systems), each of which was at a concentration of 0.5 μg/mL, were added to the chip and incubated for 1 h, allowing for formation of sandwich complexes around the target protein or cytokine.

Techniques: Binding Assay, Recombinant, Titration