Fig. 2

MELK promoted the migration and invasion of LUAD. a MELK expression in normal lung cells and lung cancer cells was analyzed by Western blot. b LUAD 95-D cells with relative lower levels of MELK were stably transfected with pCMV6-MELK-Myc-DDK (pMELK) for increasing MELK expression and the results were identified by Western blot. c The effects of MELK on migration and invasion in 95-D cells were measured by Transwell experiments. Bars indicates SD, P values represented the significant difference between 95-D_MELK and 95-D_NC, Student’s t test. d LUAD H1299 and H1975 cells with relative higher levels of MELK were transfected with siRNA duplexes of MELK for 72 h and detected by Western blot. e The effects of MELK on migration and invasion in H1299 and H1975 cells were measured by Transwell experiments. After transfected with siRNA duplex of NC or MELK for 48 h, H1299 and H1975 cells were seeded in the Transwell covered with or without Matrigel for invasion or migration assay. Bars indicates SD, P values represented the significant difference between siMELK group and siNC group, Student’s t test. f The effects of MELK on proteins involved in metastasis were detected by Western blot. H1299 and H1975 cells were transfected with siRNA duplexes of MELK for 72 h and collected for Western blot. g. The mRNA levels of those protein which were changed by MELK in protein levels. H1299 and H1975 cells were transfected with siRNA duplexes of MELK for 72 h and collected for qRT-PCR. h The interaction of MELK and those protein which were changed by MELK in protein levels. 293T cells were transfected with pNC or pMELK for 24 h and collected for Co-IP analysis. i The effects of Slug or Twist1 on MELK-inducing pathway of migration and invasion. 95-D_MELK cells were transfected with siRNA duplexes of Slug for 72 h and analyzed by Western blot. j Knockdown of Slug or Twist1 reduced the migration and invasion of 95-D with overexpression of MELK. 95-D_MELK cells were transfected with siRNA duplexes of Slug for 48 h, and the transwell experiments were performed. Bars indicates SD, P values represented the significant difference between siSlug or siTwist1 group and corresponding siNC group of 95-D_MELK cells, Student’s t test