Sc 393832

Proteins were extracted from tissue samples and cells using RIPA lysis buffer in the presence of protease and phosphatase inhibitors. After determining the protein concentration using the BCA method, 20 μg of protein was separated on 10% SDS–PAGE gel and then transferred onto PVDF membranes. After blocking with 5% non-fat milk and washing with TBST three times, the membranes were incubated with specific antibodies overnight at 4 °C. The next day, membranes were incubated with horseradish peroxidase-conjugated secondary antibodies (Beyotime, Nantong, China) for 1 h at room temperature. The ECL kit (Servicebio Technology) was used to visualize membranes. A primary antibody targeting TRIM26 (sc-393832, 1:500) was obtained from Santa Cruz Biotechnology (Dallas, TX, USA), while primary antibodies targeting RACK1 (#5432, 1:500), MEK1/2(#8727, 1:500), p-MEK1/2(#2338, 1:500), ERK1/2 (#4695, 1:500), p-ERK1/2 (#8544, 1:1000), E-cadherin (#3195, 1:1000), N-cadherin (#13116, 1:500), and vimentin (#5741, 1:500) were purchased from Cell Signaling Technology (Danvers, MA, USA).

Antibodies and reagents were acquired from designated suppliers:
TRIM26 (ab188017, Abcam), TRIM26 (27013-1-AP, Proteintech), TRIM26 (sc-393832, Santa Cruz Biotechnology), GPX4 (ab125066, Abcam), GPX4 (sc-166570, Santa Cruz Biotechnology), PLK1 (ab189139, Abcam), p-PLK1 (ab155095, Abcam), β-actin (ab8226, Abcam), K48-ubiquitin (ab140601, Abcam), K63-ubiquitin (12930, Cell Signaling Technology), anti-Flag (66008, Proteintech), anti-Myc (16286, Proteintech), p-S/T (61 G, abmart), p-S/T (612549, BD Biosciences), Anti-His (66005, Proteintech), anti-Flag (14793, Cell Signaling Technology), anti-Myc (2276, Cell Signaling Technology), Anti-His (12689, Cell Signaling Technology), anti-HA (3724, Cell Signaling Technology),
MDA (ab27642, Abcam), 4-HNE (ab48506, Abcam), MG132 (S1748, Beyotime), cycloheximide (HY12320, MedChemExpress), Chloroquine (HY-17589A, MedChemExpress), Onvansertib (HY-15828, MedChemExpress), MLN0905 (HY-15155, MedChemExpress), Recombinant Human PLK1 (ab271716, Abcam).