Anti hoxa10

Endometrial tissues or cells were collected and subjected to protein extraction, and the protein concentration was determined. Protein lysates (18–20 μg) were separated by SDS–PAGE, and PVDF membranes were used for protein transfer. The membranes were blocked with 5% milk at room temperature for 1 h. They were then incubated with anti-FHL1 (Abcam, 1:1000, ab49241), anti-HOXA10 (Santa Cruz, 1:1000, sc-271953), anti-SIRT2 (Abcam, 1:1000, ab211033), anti-FAK (Abways, 1:1000, 12363-1-AP), anti-pFAK (Bioworld, 1:1000, 103571-AP), anti-β3 Integrin (Bioworld, 1:500, BS3660), anti-Flag (Sigma, 1:10,000, A8592), anti-Myc (Invitrogen, 1:10,000, 46-0709), and anti-GAPDH (Bioworld, 1:10,000, AP0063) primary antibodies overnight at 4 °C. Specific secondary antibodies were then added, and the membrane was further incubated for 1 h at room temperature. Proteins were then detected using an enhanced chemiluminescence kit (ClarityTM Western ECL Substrate, Bio-Rad). The ratio of the corresponding GAPDH grey value was analyzed by ImageJ software, and the value was normalized as the relative protein content.

The tissue sections were deparaffinized in xylene and rehydrated in a graded series of ethanol followed by epitope retrieval in citrate buffer (pH = 6.0). HOXA10, BCL2, and Ki67 expressions were detected using antibodies anti‐HOXA10 (1:100, Santa Cruz, USA), anti‐BCL2 (1:100, CST, USA), and anti‐Ki67 (1:600; GeneTex), respectively. After the secondary antibody incubation with Envision System Plus‐HRP, the slides were rinsed and counterstained with Mayer's hematoxylin. Finally, sections were imaged and evaluated.