Anti klf4 antibody

253J B-V cells were incubated with miR-145 for 48 h and then immunostained with PKM2 antibody according to the immunofluorescence protocol of Cell Signaling Technology. The nuclei were stained with Hoechst 33342, and for actin labeling the cells were incubated with the fluorescent F-actin probe Rhodamine Phalloidin (Cytoskeleton, Denver, CO, USA). The cells were viewed with a BIOREVO fluorescence microscope (Keyence, Osaka, Japan). Standard immunohistochemical staining in clinical samples was performed using anti-KLF4 antibody (Santa Cruz Biotechnology and Proteintech Group, Inc, Rosemont, IL, USA).

Esophageal squamous cell carcinoma (ESCC) tissues from 38 patients who had received esophagectomy were collected in Cancer Hospital, Chinese Academy of Medical Science from December, 2000 to June, 2001. The follow-up period was from 3 months to 92 months, with a mean period of 39.6 months. The disease-free survival time was defined as the time between diagnosis of esophageal squamous cell carcinoma and recurrence of disease. The study was approved by the medical ethics committee of Cancer Hospital, Chinese Academy of Medical Science.
Slides were probed with anti-KLF4 antibody (1:100, Santa Cruz) and assessed by two individual investigators. KLF4 immunoreactivity was scored 0~3 by the color and percentage of cells showing distinct nuclear and/or diffuse cytoplasmic immunohistochemical reaction. Scores of 0~1 were defined as “low expression”, and scores of 2~3 as “high expression”.