Gapdh rabbit monoclonal antibody

Quercetin (Que) standard (purity ≥98%) was purchased from MedChemExpress (New Jersey, USA). Trichloromethane, methyl alcohol, and other chemicals were purchased from Sinopharm Chemical Reagent Co., Ltd. Soya bean lecithin (purity ≥98%) was purchased from Suzhou Meilun Biotechnology Co., Ltd. Cholesterol (purity ≥97%) was obtained from Accela ChemBio Co., Ltd. L‐α‐glycerophosphocholine (GPC; purity ≥98%), DSPE‐PEG2000, TPP‐DSPE‐PEG2000, FITC‐DSPE‐PEG2000, and Cy5.5‐DSPE‐PEG2000 were purchased from Xi'an Ruixi Biological Technology Co., Ltd (Shanxi, China). Dulbecco's modified eagle medium (DMEM), penicillin/streptomycin, and fetal bovine serum were obtained from Life Technologies (Carlsbad, CA, USA). We purchased 2′,7′‐dichlorofluorescin diacetate (H₂DCF‐DA), dihydroethidium (DHE), neuronal Class III β‐TUBULIN (Tuj1) monoclonal antibody, GAPDH rabbit monoclonal antibody, IL‐1β rabbit polyclonal antibody, and IBA1 rabbit polyclonal antibody from Beyotime Biotechnology Co., Ltd. (Shanghai, China). Further, 4′,6‐diamidino‐2‐phenylindole (DAPI) was purchased from Amy Jet Scientific Inc Co., Ltd. (Wuhan, China).

After transfection for 24 h, proteins were extracted from the cells using the radio immunoprecipitation assay lysis buffer (Beyotime, Nantong, China) on ice. Protein concentration was measured via the bicinchoninic acid protein assay kit (Beyotime). Equal amount of protein (20 μg) was separated by 10% sodium dodecyl sulfate–polyacrylamide gel electrophoresis and transferred onto polyvinylidene fluoride membranes, which were then blocked for an hour with 5% skimmed milk powder. Next the membranes were incubated with the primary antibodies including anti-PABPC1 (ab233280), anti-E-cadherin (ab1416), anti-N-cadherin (ab202030), anti-snail (ab229701) and anti-vimentin (ab193555) overnight at 4°C. Additionally, anti-PABPC1 and all EMT-related protein primary antibodies were purchased from Abcam Trading Company Ltd (Shanghai, China). GAPDH rabbit monoclonal antibody (AF1186; Beyotime) was regarded as an internal control. Subsequently, these membranes were washed with Tris-buffered saline Tween thrice for 5 min and were incubated with the horseradish peroxidase-conjugated secondary antibodies for 1 h at room temperature. The protein luminescence effect was evaluated via an enhanced chemiluminescence plus with a bioimaging system. Image J software served to scan the gray values.