A13685

From colon tissues and IEC6 cells, total proteins were isolated using a lysis buffer. These proteins were subjected to heat at 98 °C for a short duration of 10 min. Following this, they were electrophoretically separated on a 10% SDS-PAGE and subsequently electrotransferred onto PVDF membranes. After the transfer, the membranes were blocked with a 5% non-fat milk solution for an hour at ambient temperature. Various primary antibodies were then introduced: GPX4 (1:500, A1933, abclonal, Wuhan, China), SLC7A11 (1:500, A13685, abclonal, Wuhan, China), TFR1 (1:500, A5865, abclonal, Wuhan, China), Nrf2 (1:500, A0674, abclonal, Wuhan, China), HO-1 (1:500, A19062, abclonal, Wuhan, China), and NQO-1 (1:500, A19586, abclonal, Wuhan, China), which were incubated with the membranes overnight in a 4 °C environment. This was followed by a one-hour exposure to a secondary antibody (1:7000, AS003, abclonal, Wuhan, China). For normalization, β-actin (1:5000, AC026, abclonal, Wuhan, China) served as an internal reference protein to evaluate relative protein expression levels. The membranes’ imaging was eventually carried out, with optical densities of the bands of interest being quantified using the AlphaEaseFC software4.0.0 suite from Alpha Innotech Corp, 3040 Oakmead Village Drive, Santa Clara, CA, USA.

In a tissue microarray (TMA), 80 ovarian benign cyst tissues, 117 OC tissues, and 69 peritoneal metastatic OC tissues were assembled and analyzed with an anti-CEBPG antibody (12997-1-AP, Proteintech, 1:100 dilution) or an anti-SLC7A11 antibody (A13685, ABclonal, 1:100 dilution).