Fig 1: High-Level Amplification of FGFR1 and LETM2 Affected by SVs(A) The top panel represents different types of SVs indicated by lines with different colors on chromosome 8 in ESCC-16T; the middle panel shows normalized coverage for each window. Zoom-in view of high-level amplification of FGFR1 and LETM2 locus is shown in the bottom panel. FISH analysis demonstrates DM-derived amplification of FGFR1. Scale bar represents 10 µm.(B) High-level amplification of FGFR1 in ESCC-06T. Top: FGFR1 locus and the high-level amplification region containing MYC gene are shown. Bottom: Zoom-in view of high-level amplification of FGFR1 locus. FISH confirms CNAs by showing FGFR1 amplification as clustered multiple green signals. Scale bar represents 10 µm.(C) FISH confirms CNAs by showing LETM2 amplification as scattered multiple green signals. Scale bar represents 10 µm.(D) Immunohistochemical analysis shows LETM2 staining in ESCCs.(E) LETM2 expression level in multiple ESCC lines determined by RT-PCR and western blotting.(F) LETM2 knockdown prevents cell proliferation but has no effect on cell migration/invasion as monitored by MTT or in vitro cell migration and invasion assays in KYSE150 and ECA109 cells. Knockdown of LETM2 is demonstrated by immunoblotting; GAPDH was used as loading control.Data are mean ± SD; each experiment was performed in triplicate. **p < 0.01.