Sft2d1

We added a mixture of cell lysis buffer (Servicebio, China) and protease inhibitor PMSF to the cells. The sample was then denatured by adding SDS-loading buffer and subjected to electrophoresis. The PVDF membrane (Millipore, USA) was wet-transferred at a constant current. After blocking with skim milk at room temperature for 2 h, the primary antibody SFT2D1 (Immunoway, USA, 1:1000) was incubated at 4℃ overnight. The secondary antibody (bioworld, USA, 1:10000) was incubated at room temperature for 1 h, followed by detection with a developing solution.

We collected normal cervical tissue and cervical cancer tissue at the Hunan Provincial Cancer Hospital for immunohistochemical staining, and it has been reviewed and approved by the Ethics Committee of Hunan Cancer Hospital. After dewaxing of sections, heat antigen retrieval was performed. The primary antibodies SFT2D1 (Immunoway, USA, 1:100) and CD31 (ZenBio, China, 1:100) were incubated overnight at 4 °C. The secondary antibodies were incubated for 20 min using the PV-9000 kit (ZSGB-BIO, China). DAB reagent (ZSGB-BIO, China) was used for antibody staining, with brown-yellow indicating positive signal areas. Cell nuclei were stained blue with hematoxylin (Servicebio, China). Images were captured using microscope (Zeiss, Germany) and analyzed using Image J software (1.53, USA).