Protein Expression Analysis in MGC-803 Cells

Protein was extracted from MGC-803 cells at different groups and tumor tissues using RIPA buffer containing PMSF. Proteins (50 µg) were resolved on 10% SDS-PAGE and electrotransferred on polyvinylidene fluoride membranes (Invitrogen, CA, USA). The primary antibodies used here were anti-PI3K, anti-p-PI3K, anti-AKT, anti-p-AKT, anti-mTOR, anti-p-mTOR, anti-E-cadherin, anti-N-cadherin, anti-Vimentin, anti-p53, anti-CDK4, anti-CyclinD1 or anti-beta-actin from Cell Signaling Technology or Abcam. Then, the membranes were incubated with secondary antibody and target proteins were analyzed by ImageLab software (Bio-Rad, CA, USA) [25 (link)].

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Gao F., Li R., Wei P.F., Ou L., Li M., Bai Y., Luo W.J, & Fan Z. (2022). Synergistic anticancer effects of everolimus (RAD001) and Rhein on gastric cancer cells via phosphoinositide-3-kinase (PI3K)/protein kinase B (AKT)/mammalian target of rapamycin (mTOR) pathway. Bioengineered, 13(3), 6332-6342.

Publication 2022

polyvinylidene fluoride membranes anti-PI3K anti-p-PI3K anti-AKT anti-p-AKT anti-mTOR anti-p-mTOR anti-E-cadherin anti-N-cadherin anti-Vimentin anti-p53 anti-CDK4 anti-CyclinD1 anti-beta-actin ImageLab software

Antibodies Antibody Beta actin Buffer Cadherin Membranes Mtor Pi3k Polyvinylidene fluoride Proteins Ripa Sds page Tissues Tumor Vimentin

Corresponding Organization : Affiliated Hospital of Shaanxi University of Chinese Medicine

Other organizations : Shaanxi University of Chinese Medicine

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Variable analysis

independent variables

Groups (different groups of MGC-803 cells)
Tumor tissues

dependent variables

Expression levels of PI3K, p-PI3K, AKT, p-AKT, mTOR, p-mTOR, E-cadherin, N-cadherin, Vimentin, p53, CDK4, CyclinD1, and beta-actin

control variables

Protein amount (50 µg)
SDS-PAGE (10%)
Polyvinylidene fluoride membranes
Primary antibodies (anti-PI3K, anti-p-PI3K, anti-AKT, anti-p-AKT, anti-mTOR, anti-p-mTOR, anti-E-cadherin, anti-N-cadherin, anti-Vimentin, anti-p53, anti-CDK4, anti-CyclinD1, anti-beta-actin)
Secondary antibody
ImageLab software (Bio-Rad, CA, USA)

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