Fig. 4
From: Role of Raf-kinase inhibitor protein in colorectal cancer and its regulation by hydroxycamptothecine
Establishment of the single clone SW1116/RKIP— cells and effect of shRNA vector (PGPU6/GFP/Neo-PEBP1-501/271/376 targeted RKIP), over-expression vector (pIRES2-EGFP/PEBP-1). a SW1116 cells stably transfected with shRNA 271 were screened with G418 in 800ug/mL to obtain single colony selection in 24 well culture plates under optical microscope; b A under fluorescence microscope; c SW1116 cells were stably transfected with either empty shRNA vector PGPU6/ GFP/Neo (SW1116 con) or shRNA vector PGPU6/GFP/Neo-PEBP1 -501/271/376 target RKIP (shRNA 501, 271,376) respectively. RKIP expression was analyzed by Western Blot. shRNA 271 caused significant down-regulation of RKIP expression. d In SW1116/RKIP— cells, Western Blot verified the decrease in RKIP expression. e LoVo control: LoVo cells transfected with empty pIRES2-EGFP; PEBP-1-a: LoVo cells transfected with over-expression vector pIRES2-EGFP/PEBP-1, PEBP-1-b is repeated sample to PEBP-1-a. a, b, Magnification ×400). Abbreviations: RKIP, Raf kinase inhibitor protein; NCRCT, non neoplastic colorectal tissue; CRCT, colorectal cancer tissue; MLNT, metastatic lymph node tissue; PEBP, Phosphatidylethanolamine-binding proteins