Fig 2: Identification of the efficiencies of EFEMP1 overexpression and knockdown after transfection (S1 and S21: non-infected control, S1-NC and S21-NC: infected with control-shRNA, S1-124: infected with EFEMP1-shRNA 124, S1-125: infected with EFEMP1-shRNA 125, S1-126: infected with EFEMP1-shRNA 126, and S21-exEFEMP1: infected with pLVX- EFEMP1-Puro vector)(A) EFEMP1 protein expression in lentivirus-infected cells as measured by Western blot. (B) EFEMP1 mRNA expression in lentivirus-infected cells as measured by q-RT-PCR. (C) PCR products as measured by agarose gel electrophoresis. (D) EFEMP1 protein expression in lentivirus-infected cells as measured by ICC staining (Magnification × 200). *P < 0.05 versus control.

Fig 3: Effects of EFEMP1 on cell cycle(A) Cell cycle analysis as measured by flow cytometry in lentivirus-infected cells. (B) EFEMP1 knockdown suppressed cell by blocking their progression from the G1/G0 phase to S phase, while EFEMP1 overexpression promoted cell proliferation by increasing the S phase cells. (C) Cell cycle-related proteins Cyclin A, Cyclin B1, Cyclin D1 and Cyclin E in lentivirus-infected cells as measured by Western blot. (D) The average band intensities of Cyclin A, Cyclin B1, Cyclin D1 and Cyclin E normalized to GAPDH. *P < 0.05 versus control.

Fig 4: Effects of EFEMP1 on cell migration and invasion(A) Cell migration images of lentivirus-infected cells as measured by Boyden chambers without Matrigel. (Magnification × 200). (B) Cell invasion images of lentivirus-infected cells by Boyden chambers coated with Matrigel. (Magnification × 200). (C) EFEMP1 knockdown suppressed cell migrating abilities, while EFEMP1 overexpression promoted cell migrating abilities. (D) EFEMP1 knockdown suppressed cell invading abilities, while EFEMP1 overexpression promoted cell invading abilities. *P < 0.05 versus control.

Fig 5: Effects of EFEMP1 on cell growth and cell colony formation(A) Knockdown of EFEMP1 inhibited cell proliferation of the highly invasive subclone. (B) Overexpression of EFEMP1 promoted cell proliferation of the low invasive subclone. (C) The colony images of lentivirus-infected cells as examined by soft agar colony formation assay. (D) EFEMP1 knockdown decreased the colony forming capacity of highly invasive subclone, while overexpression of EFEMP1 increased the colony forming capacity of low invasive subclone. *P < 0.05 versus control.