figs2.

Cloning of ARPC1B gene and validation of its role in cancer metastasis. Cloning of perturbated gene using Splinkerette PCR (A). The chromosomal location of GSV insertion in M45 clone (obtained from lung metastasis) was determined by ENSEMBLE (B). The PCR product was sequenced to identify the chromosome sequence flanked by GSV (C). The insertion site of GSV was identified using UCSC genome browser (D). The identity of the gene was validated by genomic PCR (E). The reversal of the migration ability by doxycycline in M45 clone but not 2D2 cells (F). The quantification of cell migration in M45 and 2D2 clone in presence or absence of doxycycline (G). Cell models with overexpression of ARPC1A, low expression of ARPC1B, and overexpression of ARPC1A and low expression of ARPC1B were established (H). In vitro experiments confirmed that overexpression of ARPC1A and low expression of ARPC1B promote the invasion and migration ability of tumor cells (I). The quantification of cell migration in ORF-ARPC1A group, sh-ARPC1B group and ORF- ARPC1A + sh-ARPC1B(J). Correlation between ARPC1A or ARPC1B expression and survival of patients with pancreatic cancer (K, I).